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<p>Monoclonal antibody raised in mouse against amino acids 304-428 of the human <strong>GR (glucocorticoid receptor)</strong>, using a chimeric protein.</p>',
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP assays were performed using HeLa cells, the Diagenode monoclonal antibody directed against GR (Cat. No. MAb-010-050) and optimized PCR primer sets for qPCR. The cells were treated either with ethanol (EtOH, used as a negative control) or triamcinolone acetonide (TA) for 4 hours prior to cell harvesting. ChIP was performed using sheared chromatin from 3 million cells and 5 μg of antibody. QPCR was performed with primers for the human metallothionein promoter (hMTIIA) and for exon 2 of the human myoglobin gene (hmyo ex2), used as a negative control. Figure 1 shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +/- control target). These results demonstrate the occupancy of the human metallothionein IIA promoter by GR. </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP was performed on sheared chromatin from 3.5 million HeLaB2 cells using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). The cells were treated with the synthetic GR ligand triamcinolone acetonide (TA) for 4 hours prior to harvesting. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence of chromosome 16 (figure 2A) as well as the MT2A positive control gene (figure 2B). The position of the PCR amplicon is also indicated. Figure 2C, D and E show the results for the known GR target genes PER1 on chromosome 17 and FKBP5 and TNFAIP3 on chromosome 6. </small></p>
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<p><small><strong> Figure 3. Sandwich ELISA </strong><br />The specificity of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050) was assessed by sandwich ELISA. Figure 3A: schematic representation of the sandwich ELISA with the monoclonal antibody against hGR (clone #:m2F8). Figure 3B: ELISA results using the monoclonal antibody against hGR at a concentration of 0.5 μg/ml. The figure shows an ELISA signal which is proportionally increasing with increasing amounts of recombinant hGR. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody against hGR </strong><br />Figure 4A. Extracts from HeLa cells containing the indicated amounts of GR (from 30 to 240 fmol), and from 5x10e6 Raji or Molt cells were analysed by Western blot using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). Figure 4B. Western blot analysis of extracts from 300,000 HeLa cells with the Diagenode monoclonal antibody against hGR (concentration 1 μg/ml). </small></p>
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<p><small><strong> Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR </strong><br />1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml. <br />2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml. </small></p>
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<p><small><strong> Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR </strong><br />The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above </small></p>
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<p>Monoclonal antibody raised in mouse against amino acids 304-428 of the human <strong>GR (glucocorticoid receptor)</strong>, using a chimeric protein.</p>',
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP was performed on sheared chromatin from 3.5 million HeLaB2 cells using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). The cells were treated with the synthetic GR ligand triamcinolone acetonide (TA) for 4 hours prior to harvesting. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence of chromosome 16 (figure 2A) as well as the MT2A positive control gene (figure 2B). The position of the PCR amplicon is also indicated. Figure 2C, D and E show the results for the known GR target genes PER1 on chromosome 17 and FKBP5 and TNFAIP3 on chromosome 6. </small></p>
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<p><small><strong> Figure 3. Sandwich ELISA </strong><br />The specificity of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050) was assessed by sandwich ELISA. Figure 3A: schematic representation of the sandwich ELISA with the monoclonal antibody against hGR (clone #:m2F8). Figure 3B: ELISA results using the monoclonal antibody against hGR at a concentration of 0.5 μg/ml. The figure shows an ELISA signal which is proportionally increasing with increasing amounts of recombinant hGR. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody against hGR </strong><br />Figure 4A. Extracts from HeLa cells containing the indicated amounts of GR (from 30 to 240 fmol), and from 5x10e6 Raji or Molt cells were analysed by Western blot using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). Figure 4B. Western blot analysis of extracts from 300,000 HeLa cells with the Diagenode monoclonal antibody against hGR (concentration 1 μg/ml). </small></p>
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<p><small><strong> Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR </strong><br />1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml. <br />2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml. </small></p>
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<p><small><strong> Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR </strong><br />The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2A.png" alt="GR Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2B.png" alt="GR Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2C.png" alt="GR Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2D.png" alt="GR Antibody validated in ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2E.png" alt="GR Antibody ChIP-seq Grade" /></p>
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<div class="small-6 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP was performed on sheared chromatin from 3.5 million HeLaB2 cells using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). The cells were treated with the synthetic GR ligand triamcinolone acetonide (TA) for 4 hours prior to harvesting. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence of chromosome 16 (figure 2A) as well as the MT2A positive control gene (figure 2B). The position of the PCR amplicon is also indicated. Figure 2C, D and E show the results for the known GR target genes PER1 on chromosome 17 and FKBP5 and TNFAIP3 on chromosome 6. </small></p>
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<p></p>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig3.png" alt="GR Antibody ELISA Validation" /></p>
</div>
<div class="small-6 columns">
<p><small><strong> Figure 3. Sandwich ELISA </strong><br />The specificity of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050) was assessed by sandwich ELISA. Figure 3A: schematic representation of the sandwich ELISA with the monoclonal antibody against hGR (clone #:m2F8). Figure 3B: ELISA results using the monoclonal antibody against hGR at a concentration of 0.5 μg/ml. The figure shows an ELISA signal which is proportionally increasing with increasing amounts of recombinant hGR. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig4.png" alt="GR Antibody validated in Western Blot" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody against hGR </strong><br />Figure 4A. Extracts from HeLa cells containing the indicated amounts of GR (from 30 to 240 fmol), and from 5x10e6 Raji or Molt cells were analysed by Western blot using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). Figure 4B. Western blot analysis of extracts from 300,000 HeLa cells with the Diagenode monoclonal antibody against hGR (concentration 1 μg/ml). </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig5.png" alt="GR Antibody validated in Immunohistochemistry" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig6.png" alt="GR Antibody validated for Immunohistochemistry" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR </strong><br />1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml. <br />2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig7.png" alt="GR Antibody validated in Immunoprecipitation" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR </strong><br />The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above </small></p>
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'description' => '<p><a href="https://go.diagenode.com/bioruptor-upgrade"><img src="https://www.diagenode.com/img/banners/banner-br-trade.png" /></a></p>
<p>The Bioruptor® Pico (2013-2019) represented a breakthrough for shearing micro-volumes of 5 μl to larger volumes of up to 2 ml. <span>The new generation keeps the features you like the most and bring even more innovation. Check it now:</span></p>
<center><span></span></center><center><a href="https://www.diagenode.com/p/bioruptorpico2"> <img alt="New Bioruptor Pico" src="https://www.diagenode.com/img/product/shearing_technologies/new-pico-product-banner.jpg" /></a></center>
<p></p>
<p><span>Watch our short video about the Bioruptor Pico and how it can help you accomplish perfect shearing for any application including chromatin shearing, DNA shearing for NGS, unmatched DNA extraction from FFPE samples, RNA shearing, protein extraction, and much more.</span></p>
<p>
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<p></p>
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'label3' => 'Available chromatin shearing kits',
'info3' => '<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
<table style="width: 925px;">
<tbody>
<tr valign="middle">
<td style="width: 213px;"></td>
<td style="text-align: center; width: 208px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histones)</a></strong></td>
<td style="text-align: center; width: 180px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center; width: 154px;"><strong><a href="../p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center; width: 155px;"><strong><a href="../p/chromatin-shearing-plant-chip-seq-kit">Chromatin Shearing Kit (for Plant)</a></strong></td>
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<td style="width: 213px;">
<p style="text-align: left;"><strong>SDS concentration</strong></p>
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<p style="text-align: center;">< 0.1%</p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;">0.2%</p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;">1%</p>
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<td style="text-align: center; width: 155px;">
<p style="text-align: center;">0.5%</p>
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<td style="width: 213px;">
<p style="text-align: left;"><strong>Nuclei isolation</strong></p>
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<p style="text-align: center;">Yes</p>
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<td style="text-align: center; width: 180px;">
<p style="text-align: center;">Yes</p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;">No</p>
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<td style="text-align: center; width: 155px;">
<p style="text-align: center;">Yes</p>
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<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
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<td style="text-align: center; width: 208px;">
<p style="text-align: center;">100 million cells</p>
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<td style="text-align: center; width: 180px;">
<p style="text-align: center;">100 million cells</p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;">100 million cells</p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;">up to 25 g of tissue</p>
</td>
</tr>
<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
<p style="text-align: center;"><a href="https://www.diagenode.com/en/p/manual-chipmentation-kit-for-histones-24-rxns">ChIPmentation Kit for Histones</a></p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p style="text-align: center;"><a href="../p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;"><a href="../p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;"><a href="../p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant <br />ChIP-seq kit</a></p>
</td>
</tr>
</tbody>
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'description' => '<p><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
<div class="small-12 medium-3 large-3 columns">
<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
</div>
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<p>Diagenode’s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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'description' => '<p><span style="font-weight: 400;">All Diagenode’s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
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'meta_title' => 'Diagenode's selection of Antibodies is exclusively dedicated for Epigenetic Research | Diagenode',
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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'description' => '<div class="row">
<div class="small-10 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
</div>
<div class="small-2 columns"><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></div>
</div>
<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
<div class="row">
<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
<div class="small-12 medium-6 large-6 columns">
<p></p>
<p></p>
<p></p>
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<p></p>
<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'meta_description' => 'Diagenode Offers Extensively Validated ChIP-Grade Antibodies, Confirmed for their Specificity, and high level of Performance in Chromatin Immunoprecipitation ChIP',
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'url' => 'files/products/antibodies/Datasheet_hGR_C15200010.pdf',
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'description' => '<p style="text-align: justify;"><span>Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of large sets of genome-wide sequencing data. ChIP sequencing (ChIP-seq) has now become the gold standard method for chromatin studies, given the accuracy and coverage scale of the approach over other methods. Successful ChIP-seq, however, requires a higher level of experimental accuracy and consistency in all steps of ChIP than ever before. Particularly crucial is the quality of ChIP antibodies. </span></p>',
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'description' => '<p>More than in any other immuoprecipitation assays, quality antibodies are critical tools in many epigenetics experiments. Since 10 years, Diagenode has developed the most stringent quality production available on the market for antibodies exclusively focused on epigenetic uses. All our antibodies have been qualified to work in epigenetic applications.</p>',
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'name' => 'Coordinated glucocorticoid receptor and MAFB action inducestolerogenesis and epigenome remodeling in dendritic cells',
'authors' => 'Morante-Palacios Octavio et al.',
'description' => '<p>Abstract Glucocorticoids (GCs) exert potent anti-inflammatory effects in immune cells through the glucocorticoid receptor (GR). Dendritic cells (DCs), central actors for coordinating immune responses, acquire tolerogenic properties in response to GCs. Tolerogenic DCs (tolDCs) have emerged as a potential treatment for various inflammatory diseases. To date, the underlying cell type-specific regulatory mechanisms orchestrating GC-mediated acquisition of immunosuppressive properties remain poorly understood. In this study, we investigated the transcriptomic and epigenomic remodeling associated with differentiation to DCs in the presence of GCs. Our analysis demonstrates a major role of MAFB in this process, in synergy with GR. GR and MAFB both interact with methylcytosine dioxygenase TET2 and bind to genomic loci that undergo specific demethylation in tolDCs. We also show that the role of MAFB is more extensive, binding to thousands of genomic loci in tolDCs. Finally, MAFB knockdown erases the tolerogenic properties of tolDCs and reverts the specific DNA demethylation and gene upregulation. The preeminent role of MAFB is also demonstrated in vivo for myeloid cells from synovium in rheumatoid arthritis following GC treatment. Our results imply that, once directly activated by GR, MAFB plays a critical role in orchestrating the epigenomic and transcriptomic remodeling that define the tolerogenic phenotype.</p>',
'date' => '2022-01-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/34893889',
'doi' => '10.1093/nar/gkab1182',
'modified' => '2022-05-20 09:44:29',
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'id' => '3208',
'name' => 'The Role of S-Palmitoylation of the Human Glucocorticoid Receptor (hGR) in Mediating the Nongenomic Glucocorticoid Actions',
'authors' => 'Nicolaides N. et al.',
'description' => '<p>Background: Many rapid nongenomic glucocorticoid actions are mediated by membrane-bound glucocorticoid receptors (GRs). S-palmitoylation is a lipid post-translational modification that mediates the membrane localization of some steroid receptors. A highly homologous amino acid sequence (663YLCM KTLLL671) is present in the ligand-binding domain of hGRα, suggesting that hGRα might also undergo S-palmitoylation.</p>
<p>Aim: To investigate the role of the motif 663YLCMKTLLL671 in membrane localization of the hGRα and in mediating rapid nongenomic glucocorticoid signaling.</p>
<p>Methods and Results: We showed that the mutant receptors hGRαY663A, hGRαC665A and hGRαLL670/671AA, and the addition of the palmitoylation inhibitor 2-bromopalmitate did not prevent membrane localization of hGRα and co-localization with caveolin-1, and did not influence the biphasic activation of mitogen-activated protein kinase (MAPK) signaling pathway in the early time points. Finally, the hGRα was not shown to undergo S-palmitoylation.</p>
<p>Conclusions: The motif 663YLCMKTLLL671 does not play a role in membrane localization of hGRα and does not mediate the nongenomic glucocorticoid actions.</p>',
'date' => '2017-04-15',
'pmid' => 'http://www.jmolbiochem.com/index.php/JmolBiochem/article/view/199',
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(int) 2 => array(
'id' => '2923',
'name' => 'Three Novel Heterozygous Point Mutations of NR3C1 causing Glucocorticoid Resistance',
'authors' => 'Vitellius G et al.',
'description' => '<p>Generalized glucocorticoid resistance is associated with glucocorticoid receptor (GR, NR3C1) mutations. Three novel heterozygous missense NR3C1 mutations (R477S, Y478C and L672P) were identified in patients presenting with adrenal incidentalomas, glucocorticoid excess without Cushing syndrome. Dexamethasone (DXM) binding studies demonstrated that the affinity of GR<sub>R477S</sub> and GR<sub>Y478C</sub> mutants, located in the DNA-binding domain of GR, was similar to wild-type GR (Kd = 2-3 nM). In contrast, GR<sub>L672P</sub> mutant, located in the ligand-binding domain of GR, was unable to bind glucocorticoids and was more sensitive to protein degradation. GR subcellular distribution revealed a marked decrease in DXM-induced nuclear translocation of GR<sub>R477S</sub> and GR<sub>Y478C</sub> mutants, whereas GR<sub>L672P</sub> remained exclusively cytoplasmic. Chromatin immunoprecipitation demonstrated impaired recruitment of DBD mutants onto the regulatory sequence of FKBP5. Transactivation assays disclosed the lack of transcriptional activity of GR<sub>R477S</sub> and GR<sub>L672P</sub> while GR<sub>Y478C</sub> had a reduced transactivation capacity. Three-D modeling indicated that R477S lost two essential hydrogen bonds with DNA, Y478C resulted in altered interaction with surrounding amino-acids, destabilizing DBD, while L672P altered the H8 helix folding, leading to unstructured LBD. This study identifies novel NR3C1 mutations with their molecular consequences on altered GR signaling and suggests that genetic screening of NR3C1 should be conducted in patients with subclinical hypercorticism.</p>',
'date' => '2016-04-27',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/27120390',
'doi' => '10.1002/humu.23008',
'modified' => '2016-05-13 17:32:26',
'created' => '2016-05-13 17:32:26',
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'id' => '2967',
'name' => 'Functional characterization of the hGRαT556I causing Chrousos syndrome',
'authors' => 'Nicolaides NC et al.',
'description' => '<h4>BACKGROUND:</h4>
<p><abstracttext label="BACKGROUND" nlmcategory="BACKGROUND">Chrousos syndrome is a rare pathologic condition characterized by generalized, partial resistance of target tissues to glucocorticoids and caused by inactivating mutations of the human glucocorticoid receptor (hGR) gene. A novel case of Chrousos syndrome has been reported in a patient with adrenal incidentaloma, who harboured a heterozygous point mutation in the hGR gene, which resulted in threonine (T) to isoleucine (I) substitution at amino acid position 556 in the ligand-binding domain of the receptor.</abstracttext></p>
<h4>OBJECTIVE:</h4>
<p><abstracttext label="OBJECTIVE" nlmcategory="OBJECTIVE">To delineate the molecular mechanisms through which the mutant receptor hGRαT556I causes Chrousos syndrome.</abstracttext></p>
<h4>DESIGN AND RESULTS:</h4>
<p><abstracttext label="DESIGN AND RESULTS" nlmcategory="RESULTS">Compared with the wild-type receptor, the mutant receptor hGRαT556I demonstrated 50% reduction in its ability to transactivate glucocorticoid-responsive genes and in the affinity for the ligand, 30% increase in the ability to transrepress the nuclear factor-κB-target genes and a 3,4-fold delay in the cytoplasmic-to-nuclear translocation. The mutant receptor hGRαT556I did not exert a dominant negative effect upon the hGRα-mediated transcriptional activity; it preserved its ability to bind to DNA and interacted with the glucocorticoid receptor-interacting protein 1 coactivator mostly through its activation function-1 domain. Structural biology studies revealed that the T556I mutation caused disruption of the hydrogen bond formed by the T556 with the =O group of P637 backbone, which resulted in a significant relocation of the P637-bearing loop. This conformational alteration affected the local 3D arrangement of the receptor and hence the electrostatic surface of the region.</abstracttext></p>
<h4>CONCLUSIONS:</h4>
<p><abstracttext label="CONCLUSIONS" nlmcategory="CONCLUSIONS">The hGRαT556I causes Chrousos syndrome by impairing multiple steps of the glucocorticoid signal transduction pathway.</abstracttext></p>',
'date' => '2016-01-01',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/26541474',
'doi' => ' 10.1111/eci.12563',
'modified' => '2016-06-29 09:41:42',
'created' => '2016-06-29 09:41:42',
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'id' => '2939',
'name' => 'Transient generalized glucocorticoid hypersensitivity',
'authors' => 'Nicolaides NC et al.',
'description' => '<h4>BACKGROUND:</h4>
<p><abstracttext label="BACKGROUND" nlmcategory="BACKGROUND">Transient generalized glucocorticoid hypersensitivity is a rare disorder characterized by increased tissue sensitivity to glucocorticoids and compensatory hypo-activation of the hypothalamic-pituitary-adrenal axis. The condition itself and the underlying molecular mechanisms have not been elucidated.</abstracttext></p>
<h4>OBJECTIVE:</h4>
<p><abstracttext label="OBJECTIVE" nlmcategory="OBJECTIVE">To present the clinical manifestations, endocrinologic evaluation and transcriptomic profile in a patient with transient generalized glucocorticoid hypersensitivity.</abstracttext></p>
<h4>DESIGN AND RESULTS:</h4>
<p><abstracttext label="DESIGN AND RESULTS" nlmcategory="RESULTS">A 9-year-old girl presented with an 8-month history of clinical manifestations suggestive of Cushing syndrome. Endocrinologic evaluation revealed undetectable 08:00 h ACTH (<1 pg/mL) and cortisol (0·025 μg/dL) concentrations, which remained decreased throughout the 24-h period and did not respond to stimulation with ovine CRH. The disease gradually resolved spontaneously over the ensuing 3 months. Sequencing of the human glucocorticoid receptor gene revealed no mutations or polymorphisms. Western blot analysis in peripheral blood mononuclear cells revealed equal protein expression of hGRα of the patient in the disease and postresolution phases compared with a control subject. Transcriptomic analysis in peripheral blood mononuclear cells in the disease and postresolution phases identified 903 differentially expressed genes. Of these, 106 genes were up-regulated and 797 were down-regulated in the disease compared with the resolution phase. Bioinformatics analysis on the differentially expressed gene networks revealed Nuclear Factor-κB as the predominant transcription factor influencing the expression of the majority of differentially expressed genes.</abstracttext></p>
<h4>CONCLUSIONS:</h4>
<p><abstracttext label="CONCLUSIONS" nlmcategory="CONCLUSIONS">Our findings indicate that a transient postreceptor defect, or a virus- or bacterium-encoded molecule, may have enhanced glucocorticoid signal transduction, leading to transient generalized glucocorticoid hypersensitivity and hypo-activation of the HPA axis.</abstracttext></p>',
'date' => '2015-12-01',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/26479047',
'doi' => ' 10.1111/eci.12554',
'modified' => '2016-05-27 10:41:26',
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'description' => 'Bisphenol A (BPA) is an estrogen-mimicking endocrine disruptor. Early-life exposures to low doses of BPA exert long-lasting effects on animals' reproductive and brain physiology. However, little is known about the effects of BPA on the stress-response system. Given the interaction of sex and stress hormones, we examined the effect of a low perinatal BPA exposure on the function of the hypothalamic-pituitary-adrenal (HPA) axis at rest and upon application of acute stress. Throughout pregnancy and lactation rats received daily 40 μg BPA/kg body weight orally via cornflakes. We studied the effect of this low but chronic exposure to BPA in the male and female offspring at puberty. BPA exposure led to abnormal adrenal histology including reduced zona reticularis especially in male offspring, hyperplasia of zona fasciculata in both sexes, and increased adrenal weight in female offspring. BPA-treated females had increased basal corticosterone and reduced hypothalamic glucocorticoid receptors (GR) levels. Stressed BPA-exposed females exhibited anxiety-like behavioral coping, a less rigorous corticosterone response, and did not downregulate GR in the hypothalamus, compared with control females. BPA-exposed males exhibited a heightened corticosterone stress response compared with females; they also displayed increased pro-opiomelanocortin mRNA levels and retained the prestress levels of pituitary corticotropin-releasing hormone-receptor 1, compared with control males. We found that perinatal chronic exposure to a low dose of BPA perturbs the basal and stress-induced activity of the HPA axis in a sexually dimorphic manner at adolescence. Exposure to BPA might contribute to increased susceptibility to stress-related disorders in later life.',
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'description' => 'Glucocorticoid receptor (GR) exerts anti-inflammatory action in part by antagonizing proinflammatory transcription factors such as the nuclear factor kappa-b (NFKB). Here, we assess the crosstalk of activated GR and RELA (p65, major NFKB component) by global identification of their binding sites and target genes. We show that coactivation of GR and p65 alters the repertoire of regulated genes and results in their association with novel sites in a mutually dependent manner. These novel sites predominantly cluster with p65 target genes that are antagonized by activated GR and vice versa. Our data show that coactivation of GR and NFKB alters signaling pathways that are regulated by each factor separately and provide insight into the networks underlying the GR and NFKB crosstalk.',
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<p>The Bioruptor® Pico (2013-2019) represented a breakthrough for shearing micro-volumes of 5 μl to larger volumes of up to 2 ml. <span>The new generation keeps the features you like the most and bring even more innovation. Check it now:</span></p>
<center><span></span></center><center><a href="https://www.diagenode.com/p/bioruptorpico2"> <img alt="New Bioruptor Pico" src="https://www.diagenode.com/img/product/shearing_technologies/new-pico-product-banner.jpg" /></a></center>
<p></p>
<p><span>Watch our short video about the Bioruptor Pico and how it can help you accomplish perfect shearing for any application including chromatin shearing, DNA shearing for NGS, unmatched DNA extraction from FFPE samples, RNA shearing, protein extraction, and much more.</span></p>
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'label2' => 'Recommended settings for DNA shearing with Bioruptor® Pico',
'info2' => '<p>Follow our guidelines and find the good parameters for your expected DNA size: <a href="https://pybrevet.typeform.com/to/o8cQfM">DNA shearing with the Bioruptor® Pico</a></p>
<p></p>
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'label3' => 'Available chromatin shearing kits',
'info3' => '<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
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<td style="text-align: center; width: 208px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histones)</a></strong></td>
<td style="text-align: center; width: 180px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center; width: 154px;"><strong><a href="../p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center; width: 155px;"><strong><a href="../p/chromatin-shearing-plant-chip-seq-kit">Chromatin Shearing Kit (for Plant)</a></strong></td>
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<p style="text-align: center;">< 0.1%</p>
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<td style="text-align: center; width: 180px;">
<p style="text-align: center;">0.2%</p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;">1%</p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;">0.5%</p>
</td>
</tr>
<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Nuclei isolation</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;">Yes</p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;">Yes</p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;">No</p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;">Yes</p>
</td>
</tr>
<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Allows for shearing of... cells/tissue</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;">100 million cells</p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;">100 million cells</p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;">100 million cells</p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;">up to 25 g of tissue</p>
</td>
</tr>
<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
<p style="text-align: center;"><a href="https://www.diagenode.com/en/p/manual-chipmentation-kit-for-histones-24-rxns">ChIPmentation Kit for Histones</a></p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p style="text-align: center;"><a href="../p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;"><a href="../p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;"><a href="../p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant <br />ChIP-seq kit</a></p>
</td>
</tr>
</tbody>
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'description' => 'Glucocorticoid receptor (GR) exerts anti-inflammatory action in part by antagonizing proinflammatory transcription factors such as the nuclear factor kappa-b (NFKB). Here, we assess the crosstalk of activated GR and RELA (p65, major NFKB component) by global identification of their binding sites and target genes. We show that coactivation of GR and p65 alters the repertoire of regulated genes and results in their association with novel sites in a mutually dependent manner. These novel sites predominantly cluster with p65 target genes that are antagonized by activated GR and vice versa. Our data show that coactivation of GR and NFKB alters signaling pathways that are regulated by each factor separately and provide insight into the networks underlying the GR and NFKB crosstalk.',
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'description' => '<p>Alternative names: <strong>NR3C1</strong>,<strong> GCCR</strong>, <strong>GCR</strong>, <strong>GRL</strong></p>
<p>Monoclonal antibody raised in mouse against amino acids 304-428 of the human <strong>GR (glucocorticoid receptor)</strong>, using a chimeric protein.</p>',
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP assays were performed using HeLa cells, the Diagenode monoclonal antibody directed against GR (Cat. No. MAb-010-050) and optimized PCR primer sets for qPCR. The cells were treated either with ethanol (EtOH, used as a negative control) or triamcinolone acetonide (TA) for 4 hours prior to cell harvesting. ChIP was performed using sheared chromatin from 3 million cells and 5 μg of antibody. QPCR was performed with primers for the human metallothionein promoter (hMTIIA) and for exon 2 of the human myoglobin gene (hmyo ex2), used as a negative control. Figure 1 shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +/- control target). These results demonstrate the occupancy of the human metallothionein IIA promoter by GR. </small></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2A.png" alt="GR Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2B.png" alt="GR Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2C.png" alt="GR Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2D.png" alt="GR Antibody validated in ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2E.png" alt="GR Antibody ChIP-seq Grade" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP was performed on sheared chromatin from 3.5 million HeLaB2 cells using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). The cells were treated with the synthetic GR ligand triamcinolone acetonide (TA) for 4 hours prior to harvesting. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence of chromosome 16 (figure 2A) as well as the MT2A positive control gene (figure 2B). The position of the PCR amplicon is also indicated. Figure 2C, D and E show the results for the known GR target genes PER1 on chromosome 17 and FKBP5 and TNFAIP3 on chromosome 6. </small></p>
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<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig3.png" alt="GR Antibody ELISA Validation" /></p>
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<div class="small-6 columns">
<p><small><strong> Figure 3. Sandwich ELISA </strong><br />The specificity of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050) was assessed by sandwich ELISA. Figure 3A: schematic representation of the sandwich ELISA with the monoclonal antibody against hGR (clone #:m2F8). Figure 3B: ELISA results using the monoclonal antibody against hGR at a concentration of 0.5 μg/ml. The figure shows an ELISA signal which is proportionally increasing with increasing amounts of recombinant hGR. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig4.png" alt="GR Antibody validated in Western Blot" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody against hGR </strong><br />Figure 4A. Extracts from HeLa cells containing the indicated amounts of GR (from 30 to 240 fmol), and from 5x10e6 Raji or Molt cells were analysed by Western blot using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). Figure 4B. Western blot analysis of extracts from 300,000 HeLa cells with the Diagenode monoclonal antibody against hGR (concentration 1 μg/ml). </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig5.png" alt="GR Antibody validated in Immunohistochemistry" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig6.png" alt="GR Antibody validated for Immunohistochemistry" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR </strong><br />1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml. <br />2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig7.png" alt="GR Antibody validated in Immunoprecipitation" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR </strong><br />The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above </small></p>
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<p><small><strong> Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR </strong><br />The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above </small></p>
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<div class="small-4 columns">
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<p><small><strong> Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR </strong><br />1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml. <br />2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml. </small></p>
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<p>The Bioruptor® Pico (2013-2019) represented a breakthrough for shearing micro-volumes of 5 μl to larger volumes of up to 2 ml. <span>The new generation keeps the features you like the most and bring even more innovation. Check it now:</span></p>
<center><span></span></center><center><a href="https://www.diagenode.com/p/bioruptorpico2"> <img alt="New Bioruptor Pico" src="https://www.diagenode.com/img/product/shearing_technologies/new-pico-product-banner.jpg" /></a></center>
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<p><span>Watch our short video about the Bioruptor Pico and how it can help you accomplish perfect shearing for any application including chromatin shearing, DNA shearing for NGS, unmatched DNA extraction from FFPE samples, RNA shearing, protein extraction, and much more.</span></p>
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<p></p>
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'info3' => '<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
<table style="width: 925px;">
<tbody>
<tr valign="middle">
<td style="width: 213px;"></td>
<td style="text-align: center; width: 208px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histones)</a></strong></td>
<td style="text-align: center; width: 180px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center; width: 154px;"><strong><a href="../p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center; width: 155px;"><strong><a href="../p/chromatin-shearing-plant-chip-seq-kit">Chromatin Shearing Kit (for Plant)</a></strong></td>
</tr>
<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>SDS concentration</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;">< 0.1%</p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;">0.2%</p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;">1%</p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;">0.5%</p>
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<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Nuclei isolation</strong></p>
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<td style="text-align: center; width: 208px;">
<p style="text-align: center;">Yes</p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;">Yes</p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;">No</p>
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<td style="text-align: center; width: 155px;">
<p style="text-align: center;">Yes</p>
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<td style="width: 213px;">
<p style="text-align: left;"><strong>Allows for shearing of... cells/tissue</strong></p>
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<td style="text-align: center; width: 208px;">
<p style="text-align: center;">100 million cells</p>
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<td style="text-align: center; width: 180px;">
<p style="text-align: center;">100 million cells</p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;">100 million cells</p>
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<td style="text-align: center; width: 155px;">
<p style="text-align: center;">up to 25 g of tissue</p>
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</tr>
<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
<p style="text-align: center;"><a href="https://www.diagenode.com/en/p/manual-chipmentation-kit-for-histones-24-rxns">ChIPmentation Kit for Histones</a></p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p style="text-align: center;"><a href="../p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;"><a href="../p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;"><a href="../p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant <br />ChIP-seq kit</a></p>
</td>
</tr>
</tbody>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
<div class="small-12 medium-3 large-3 columns">
<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
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'meta_title' => 'Diagenode's selection of Antibodies is exclusively dedicated for Epigenetic Research | Diagenode',
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'description' => '<p><span style="font-weight: 400;">The list of Diagenode’s highly specific antibodies for transcription studies includes the antibodies against many transcription factors and nuclear receptors. Check the list below to see our targets.</span></p>
<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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'description' => '<div class="row">
<div class="small-10 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
</div>
<div class="small-2 columns"><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></div>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
<div class="row">
<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
<div class="small-12 medium-6 large-6 columns">
<p></p>
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<p></p>
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<p></p>
<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'description' => '<p style="text-align: justify;"><span>Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of large sets of genome-wide sequencing data. ChIP sequencing (ChIP-seq) has now become the gold standard method for chromatin studies, given the accuracy and coverage scale of the approach over other methods. Successful ChIP-seq, however, requires a higher level of experimental accuracy and consistency in all steps of ChIP than ever before. Particularly crucial is the quality of ChIP antibodies. </span></p>',
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'name' => 'Coordinated glucocorticoid receptor and MAFB action inducestolerogenesis and epigenome remodeling in dendritic cells',
'authors' => 'Morante-Palacios Octavio et al.',
'description' => '<p>Abstract Glucocorticoids (GCs) exert potent anti-inflammatory effects in immune cells through the glucocorticoid receptor (GR). Dendritic cells (DCs), central actors for coordinating immune responses, acquire tolerogenic properties in response to GCs. Tolerogenic DCs (tolDCs) have emerged as a potential treatment for various inflammatory diseases. To date, the underlying cell type-specific regulatory mechanisms orchestrating GC-mediated acquisition of immunosuppressive properties remain poorly understood. In this study, we investigated the transcriptomic and epigenomic remodeling associated with differentiation to DCs in the presence of GCs. Our analysis demonstrates a major role of MAFB in this process, in synergy with GR. GR and MAFB both interact with methylcytosine dioxygenase TET2 and bind to genomic loci that undergo specific demethylation in tolDCs. We also show that the role of MAFB is more extensive, binding to thousands of genomic loci in tolDCs. Finally, MAFB knockdown erases the tolerogenic properties of tolDCs and reverts the specific DNA demethylation and gene upregulation. The preeminent role of MAFB is also demonstrated in vivo for myeloid cells from synovium in rheumatoid arthritis following GC treatment. Our results imply that, once directly activated by GR, MAFB plays a critical role in orchestrating the epigenomic and transcriptomic remodeling that define the tolerogenic phenotype.</p>',
'date' => '2022-01-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/34893889',
'doi' => '10.1093/nar/gkab1182',
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'name' => 'The Role of S-Palmitoylation of the Human Glucocorticoid Receptor (hGR) in Mediating the Nongenomic Glucocorticoid Actions',
'authors' => 'Nicolaides N. et al.',
'description' => '<p>Background: Many rapid nongenomic glucocorticoid actions are mediated by membrane-bound glucocorticoid receptors (GRs). S-palmitoylation is a lipid post-translational modification that mediates the membrane localization of some steroid receptors. A highly homologous amino acid sequence (663YLCM KTLLL671) is present in the ligand-binding domain of hGRα, suggesting that hGRα might also undergo S-palmitoylation.</p>
<p>Aim: To investigate the role of the motif 663YLCMKTLLL671 in membrane localization of the hGRα and in mediating rapid nongenomic glucocorticoid signaling.</p>
<p>Methods and Results: We showed that the mutant receptors hGRαY663A, hGRαC665A and hGRαLL670/671AA, and the addition of the palmitoylation inhibitor 2-bromopalmitate did not prevent membrane localization of hGRα and co-localization with caveolin-1, and did not influence the biphasic activation of mitogen-activated protein kinase (MAPK) signaling pathway in the early time points. Finally, the hGRα was not shown to undergo S-palmitoylation.</p>
<p>Conclusions: The motif 663YLCMKTLLL671 does not play a role in membrane localization of hGRα and does not mediate the nongenomic glucocorticoid actions.</p>',
'date' => '2017-04-15',
'pmid' => 'http://www.jmolbiochem.com/index.php/JmolBiochem/article/view/199',
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'name' => 'Three Novel Heterozygous Point Mutations of NR3C1 causing Glucocorticoid Resistance',
'authors' => 'Vitellius G et al.',
'description' => '<p>Generalized glucocorticoid resistance is associated with glucocorticoid receptor (GR, NR3C1) mutations. Three novel heterozygous missense NR3C1 mutations (R477S, Y478C and L672P) were identified in patients presenting with adrenal incidentalomas, glucocorticoid excess without Cushing syndrome. Dexamethasone (DXM) binding studies demonstrated that the affinity of GR<sub>R477S</sub> and GR<sub>Y478C</sub> mutants, located in the DNA-binding domain of GR, was similar to wild-type GR (Kd = 2-3 nM). In contrast, GR<sub>L672P</sub> mutant, located in the ligand-binding domain of GR, was unable to bind glucocorticoids and was more sensitive to protein degradation. GR subcellular distribution revealed a marked decrease in DXM-induced nuclear translocation of GR<sub>R477S</sub> and GR<sub>Y478C</sub> mutants, whereas GR<sub>L672P</sub> remained exclusively cytoplasmic. Chromatin immunoprecipitation demonstrated impaired recruitment of DBD mutants onto the regulatory sequence of FKBP5. Transactivation assays disclosed the lack of transcriptional activity of GR<sub>R477S</sub> and GR<sub>L672P</sub> while GR<sub>Y478C</sub> had a reduced transactivation capacity. Three-D modeling indicated that R477S lost two essential hydrogen bonds with DNA, Y478C resulted in altered interaction with surrounding amino-acids, destabilizing DBD, while L672P altered the H8 helix folding, leading to unstructured LBD. This study identifies novel NR3C1 mutations with their molecular consequences on altered GR signaling and suggests that genetic screening of NR3C1 should be conducted in patients with subclinical hypercorticism.</p>',
'date' => '2016-04-27',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/27120390',
'doi' => '10.1002/humu.23008',
'modified' => '2016-05-13 17:32:26',
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'id' => '2967',
'name' => 'Functional characterization of the hGRαT556I causing Chrousos syndrome',
'authors' => 'Nicolaides NC et al.',
'description' => '<h4>BACKGROUND:</h4>
<p><abstracttext label="BACKGROUND" nlmcategory="BACKGROUND">Chrousos syndrome is a rare pathologic condition characterized by generalized, partial resistance of target tissues to glucocorticoids and caused by inactivating mutations of the human glucocorticoid receptor (hGR) gene. A novel case of Chrousos syndrome has been reported in a patient with adrenal incidentaloma, who harboured a heterozygous point mutation in the hGR gene, which resulted in threonine (T) to isoleucine (I) substitution at amino acid position 556 in the ligand-binding domain of the receptor.</abstracttext></p>
<h4>OBJECTIVE:</h4>
<p><abstracttext label="OBJECTIVE" nlmcategory="OBJECTIVE">To delineate the molecular mechanisms through which the mutant receptor hGRαT556I causes Chrousos syndrome.</abstracttext></p>
<h4>DESIGN AND RESULTS:</h4>
<p><abstracttext label="DESIGN AND RESULTS" nlmcategory="RESULTS">Compared with the wild-type receptor, the mutant receptor hGRαT556I demonstrated 50% reduction in its ability to transactivate glucocorticoid-responsive genes and in the affinity for the ligand, 30% increase in the ability to transrepress the nuclear factor-κB-target genes and a 3,4-fold delay in the cytoplasmic-to-nuclear translocation. The mutant receptor hGRαT556I did not exert a dominant negative effect upon the hGRα-mediated transcriptional activity; it preserved its ability to bind to DNA and interacted with the glucocorticoid receptor-interacting protein 1 coactivator mostly through its activation function-1 domain. Structural biology studies revealed that the T556I mutation caused disruption of the hydrogen bond formed by the T556 with the =O group of P637 backbone, which resulted in a significant relocation of the P637-bearing loop. This conformational alteration affected the local 3D arrangement of the receptor and hence the electrostatic surface of the region.</abstracttext></p>
<h4>CONCLUSIONS:</h4>
<p><abstracttext label="CONCLUSIONS" nlmcategory="CONCLUSIONS">The hGRαT556I causes Chrousos syndrome by impairing multiple steps of the glucocorticoid signal transduction pathway.</abstracttext></p>',
'date' => '2016-01-01',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/26541474',
'doi' => ' 10.1111/eci.12563',
'modified' => '2016-06-29 09:41:42',
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'id' => '2939',
'name' => 'Transient generalized glucocorticoid hypersensitivity',
'authors' => 'Nicolaides NC et al.',
'description' => '<h4>BACKGROUND:</h4>
<p><abstracttext label="BACKGROUND" nlmcategory="BACKGROUND">Transient generalized glucocorticoid hypersensitivity is a rare disorder characterized by increased tissue sensitivity to glucocorticoids and compensatory hypo-activation of the hypothalamic-pituitary-adrenal axis. The condition itself and the underlying molecular mechanisms have not been elucidated.</abstracttext></p>
<h4>OBJECTIVE:</h4>
<p><abstracttext label="OBJECTIVE" nlmcategory="OBJECTIVE">To present the clinical manifestations, endocrinologic evaluation and transcriptomic profile in a patient with transient generalized glucocorticoid hypersensitivity.</abstracttext></p>
<h4>DESIGN AND RESULTS:</h4>
<p><abstracttext label="DESIGN AND RESULTS" nlmcategory="RESULTS">A 9-year-old girl presented with an 8-month history of clinical manifestations suggestive of Cushing syndrome. Endocrinologic evaluation revealed undetectable 08:00 h ACTH (<1 pg/mL) and cortisol (0·025 μg/dL) concentrations, which remained decreased throughout the 24-h period and did not respond to stimulation with ovine CRH. The disease gradually resolved spontaneously over the ensuing 3 months. Sequencing of the human glucocorticoid receptor gene revealed no mutations or polymorphisms. Western blot analysis in peripheral blood mononuclear cells revealed equal protein expression of hGRα of the patient in the disease and postresolution phases compared with a control subject. Transcriptomic analysis in peripheral blood mononuclear cells in the disease and postresolution phases identified 903 differentially expressed genes. Of these, 106 genes were up-regulated and 797 were down-regulated in the disease compared with the resolution phase. Bioinformatics analysis on the differentially expressed gene networks revealed Nuclear Factor-κB as the predominant transcription factor influencing the expression of the majority of differentially expressed genes.</abstracttext></p>
<h4>CONCLUSIONS:</h4>
<p><abstracttext label="CONCLUSIONS" nlmcategory="CONCLUSIONS">Our findings indicate that a transient postreceptor defect, or a virus- or bacterium-encoded molecule, may have enhanced glucocorticoid signal transduction, leading to transient generalized glucocorticoid hypersensitivity and hypo-activation of the HPA axis.</abstracttext></p>',
'date' => '2015-12-01',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/26479047',
'doi' => ' 10.1111/eci.12554',
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'name' => 'Perinatal exposure to low-dose bisphenol A affects the neuroendocrine stress response in rats.',
'authors' => 'Panagiotidou E, Zerva S, Mitsiou DJ, Alexis MN, Kitraki E',
'description' => 'Bisphenol A (BPA) is an estrogen-mimicking endocrine disruptor. Early-life exposures to low doses of BPA exert long-lasting effects on animals' reproductive and brain physiology. However, little is known about the effects of BPA on the stress-response system. Given the interaction of sex and stress hormones, we examined the effect of a low perinatal BPA exposure on the function of the hypothalamic-pituitary-adrenal (HPA) axis at rest and upon application of acute stress. Throughout pregnancy and lactation rats received daily 40 μg BPA/kg body weight orally via cornflakes. We studied the effect of this low but chronic exposure to BPA in the male and female offspring at puberty. BPA exposure led to abnormal adrenal histology including reduced zona reticularis especially in male offspring, hyperplasia of zona fasciculata in both sexes, and increased adrenal weight in female offspring. BPA-treated females had increased basal corticosterone and reduced hypothalamic glucocorticoid receptors (GR) levels. Stressed BPA-exposed females exhibited anxiety-like behavioral coping, a less rigorous corticosterone response, and did not downregulate GR in the hypothalamus, compared with control females. BPA-exposed males exhibited a heightened corticosterone stress response compared with females; they also displayed increased pro-opiomelanocortin mRNA levels and retained the prestress levels of pituitary corticotropin-releasing hormone-receptor 1, compared with control males. We found that perinatal chronic exposure to a low dose of BPA perturbs the basal and stress-induced activity of the HPA axis in a sexually dimorphic manner at adolescence. Exposure to BPA might contribute to increased susceptibility to stress-related disorders in later life.',
'date' => '2014-03-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/24323913',
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'authors' => 'Bei ES, Salpeas V, Alevizos B, Anagnostara C, Pappa D, Moutsatsou P',
'description' => 'Bipolar disorder (BD), a stress-related disease, is characterized by altered glucocorticoid receptor (GR) signalling. Stress response includes activation of heat shock factor (HSF) and subsequent heat shock protein (HSP) synthesis which regulate GR folding and function. The objective of this study was to investigate the possible role of HSFs, HSPs and their interaction with GR in BD. We applied immunoprecipitation, SDS-PAGE/Western blot analysis and electrophoretic mobility shift assay (EMSA) in lymphocytes (whole cell or nuclear extracts) from BD patients and healthy subjects and determined the HSPs (HSP90 and HSP70), the heterocomplexes HSP90-GR and HSP70-GR, the HSFs (HSF1 and HSF4) as well as the HSF-DNA binding. The HSP70-GR heterocomplex was elevated (p < 0.05) in BD patients vs healthy subjects, and nuclear HSP70 was reduced (p ≤ 0.01) in bipolar manic patients. Protein levels of HSF1, HSF4, HSP90, HSP90-GR heterocomplex, and HSF-DNA binding remained unaltered in BD patients vs healthy subjects. The corresponding effect sizes (ES) indicated a large ES for HSP70-GR, HSP70, HSF-DNA binding and HSF4, and a medium ES for HSP90, HSF1 and HSP90-GR between healthy subjects and bipolar patients. Significant correlations among HSFs, HSPs, GR and HSP70-GR heterocomplex were observed in healthy subjects, which were abrogated in bipolar patients. The higher interaction between GR and HSP70 and the disturbances in the relations among heat shock response parameters and GR as observed in our BD patients may provide novel insights into the contribution of these factors in BD aetiopathogenesis.',
'date' => '2013-11-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/23938235',
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'description' => 'Glucocorticoid receptor (GR) exerts anti-inflammatory action in part by antagonizing proinflammatory transcription factors such as the nuclear factor kappa-b (NFKB). Here, we assess the crosstalk of activated GR and RELA (p65, major NFKB component) by global identification of their binding sites and target genes. We show that coactivation of GR and p65 alters the repertoire of regulated genes and results in their association with novel sites in a mutually dependent manner. These novel sites predominantly cluster with p65 target genes that are antagonized by activated GR and vice versa. Our data show that coactivation of GR and NFKB alters signaling pathways that are regulated by each factor separately and provide insight into the networks underlying the GR and NFKB crosstalk.',
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<p>The Bioruptor® Pico (2013-2019) represented a breakthrough for shearing micro-volumes of 5 μl to larger volumes of up to 2 ml. <span>The new generation keeps the features you like the most and bring even more innovation. Check it now:</span></p>
<center><span></span></center><center><a href="https://www.diagenode.com/p/bioruptorpico2"> <img alt="New Bioruptor Pico" src="https://www.diagenode.com/img/product/shearing_technologies/new-pico-product-banner.jpg" /></a></center>
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'info3' => '<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
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<td style="text-align: center; width: 208px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histones)</a></strong></td>
<td style="text-align: center; width: 180px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center; width: 154px;"><strong><a href="../p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center; width: 155px;"><strong><a href="../p/chromatin-shearing-plant-chip-seq-kit">Chromatin Shearing Kit (for Plant)</a></strong></td>
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<p style="text-align: center;">< 0.1%</p>
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<p style="text-align: center;">0.2%</p>
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<td style="text-align: center; width: 154px;">
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<p style="text-align: center;">Yes</p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;">No</p>
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<p style="text-align: center;">Yes</p>
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<p style="text-align: center;">100 million cells</p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;">100 million cells</p>
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<p style="text-align: center;">up to 25 g of tissue</p>
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<p style="text-align: left;"><strong>Corresponding to shearing buffers from</strong></p>
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<td style="text-align: center; width: 208px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
<p style="text-align: center;"><a href="https://www.diagenode.com/en/p/manual-chipmentation-kit-for-histones-24-rxns">ChIPmentation Kit for Histones</a></p>
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<td style="text-align: center; width: 180px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p style="text-align: center;"><a href="../p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;"><a href="../p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;"><a href="../p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant <br />ChIP-seq kit</a></p>
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'description' => 'Glucocorticoid receptor (GR) exerts anti-inflammatory action in part by antagonizing proinflammatory transcription factors such as the nuclear factor kappa-b (NFKB). Here, we assess the crosstalk of activated GR and RELA (p65, major NFKB component) by global identification of their binding sites and target genes. We show that coactivation of GR and p65 alters the repertoire of regulated genes and results in their association with novel sites in a mutually dependent manner. These novel sites predominantly cluster with p65 target genes that are antagonized by activated GR and vice versa. Our data show that coactivation of GR and NFKB alters signaling pathways that are regulated by each factor separately and provide insight into the networks underlying the GR and NFKB crosstalk.',
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP assays were performed using HeLa cells, the Diagenode monoclonal antibody directed against GR (Cat. No. MAb-010-050) and optimized PCR primer sets for qPCR. The cells were treated either with ethanol (EtOH, used as a negative control) or triamcinolone acetonide (TA) for 4 hours prior to cell harvesting. ChIP was performed using sheared chromatin from 3 million cells and 5 μg of antibody. QPCR was performed with primers for the human metallothionein promoter (hMTIIA) and for exon 2 of the human myoglobin gene (hmyo ex2), used as a negative control. Figure 1 shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +/- control target). These results demonstrate the occupancy of the human metallothionein IIA promoter by GR. </small></p>
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<p><small><strong> Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR </strong><br />1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml. <br />2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml. </small></p>
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<p><small><strong> Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR </strong><br />The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig5.png" alt="GR Antibody validated in Immunohistochemistry" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig6.png" alt="GR Antibody validated for Immunohistochemistry" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR </strong><br />1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml. <br />2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig7.png" alt="GR Antibody validated in Immunoprecipitation" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR </strong><br />The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above </small></p>
</div>
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'info2' => '<p>Monoclonal antibody raised in mouse against amino acids 304-428 of the human GR (glucocorticoid receptor), using a chimeric protein.</p>',
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'catalog_number' => 'C15200010-50',
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'sf_code' => 'C15200010-D001-000581',
'type' => 'FRE',
'search_order' => '03-Antibody',
'price_EUR' => '360',
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'price_GBP' => '320',
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'meta_title' => 'GR Antibody - ChIP-seq Grade (C15200010) | Diagenode',
'meta_keywords' => '',
'meta_description' => 'GR (Glucocorticoid receptor) Monoclonal Antibody validated in ChIP-seq, ChIP-qPCR, WB, Flow Cyt, IP and ELISA. Batch-specific data available on the website. Alternative names: NR3C1, GCCR, GCR, GRL. Sample size avaialble.',
'modified' => '2022-01-05 14:49:18',
'created' => '2015-06-29 14:08:20',
'locale' => 'eng'
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'name' => 'GR monoclonal antibody',
'description' => 'Monoclonal antibody raised in mouse against amino acids 304-428 of the human GR (glucocorticoid receptor), using a chimeric protein.',
'clonality' => '',
'isotype' => '',
'lot' => 'NR-010',
'concentration' => '1.0 µg/µl',
'reactivity' => 'Human, rat',
'type' => 'Monoclonal',
'purity' => 'Ammonium sulphate purified',
'classification' => 'Classic',
'application_table' => '<table>
<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>ChIP/ChIP-seq</td>
<td>5 μg/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td>ELISA</td>
<td>0.5 μg/ml</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Western Blotting</td>
<td>1 μg/ml</td>
<td>Fig 4</td>
</tr>
<tr>
<td>Gel Supershift</td>
<td>5 μg/ml</td>
<td></td>
</tr>
<tr>
<td>Immunochemistry</td>
<td>2.5 μg/ml</td>
<td>Fig 5, 6</td>
</tr>
<tr>
<td>Flow cytometry</td>
<td>0.5 μg/ml</td>
<td></td>
</tr>
<tr>
<td>Immunoprecipitation</td>
<td>5 μg/IP</td>
<td>Fig 7</td>
</tr>
</tbody>
</table>
<p></p>',
'storage_conditions' => '',
'storage_buffer' => '',
'precautions' => 'This product is for research use only. Not for use in diagnostic or therapeutic procedures.',
'uniprot_acc' => '',
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'meta_keywords' => '',
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'modified' => '2020-09-18 16:34:22',
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'Master' => array(
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'name' => 'GR Antibody',
'description' => '<p>Alternative names: <strong>NR3C1</strong>,<strong> GCCR</strong>, <strong>GCR</strong>, <strong>GRL</strong></p>
<p>Monoclonal antibody raised in mouse against amino acids 304-428 of the human <strong>GR (glucocorticoid receptor)</strong>, using a chimeric protein.</p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig1.png" alt="GR Antibody ChIP Grade" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP assays were performed using HeLa cells, the Diagenode monoclonal antibody directed against GR (Cat. No. MAb-010-050) and optimized PCR primer sets for qPCR. The cells were treated either with ethanol (EtOH, used as a negative control) or triamcinolone acetonide (TA) for 4 hours prior to cell harvesting. ChIP was performed using sheared chromatin from 3 million cells and 5 μg of antibody. QPCR was performed with primers for the human metallothionein promoter (hMTIIA) and for exon 2 of the human myoglobin gene (hmyo ex2), used as a negative control. Figure 1 shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +/- control target). These results demonstrate the occupancy of the human metallothionein IIA promoter by GR. </small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2A.png" alt="GR Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2B.png" alt="GR Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2C.png" alt="GR Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2D.png" alt="GR Antibody validated in ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2E.png" alt="GR Antibody ChIP-seq Grade" /></p>
</div>
<div class="small-6 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP was performed on sheared chromatin from 3.5 million HeLaB2 cells using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). The cells were treated with the synthetic GR ligand triamcinolone acetonide (TA) for 4 hours prior to harvesting. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence of chromosome 16 (figure 2A) as well as the MT2A positive control gene (figure 2B). The position of the PCR amplicon is also indicated. Figure 2C, D and E show the results for the known GR target genes PER1 on chromosome 17 and FKBP5 and TNFAIP3 on chromosome 6. </small></p>
</div>
</div>
<p></p>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig3.png" alt="GR Antibody ELISA Validation" /></p>
</div>
<div class="small-6 columns">
<p><small><strong> Figure 3. Sandwich ELISA </strong><br />The specificity of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050) was assessed by sandwich ELISA. Figure 3A: schematic representation of the sandwich ELISA with the monoclonal antibody against hGR (clone #:m2F8). Figure 3B: ELISA results using the monoclonal antibody against hGR at a concentration of 0.5 μg/ml. The figure shows an ELISA signal which is proportionally increasing with increasing amounts of recombinant hGR. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig4.png" alt="GR Antibody validated in Western Blot" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody against hGR </strong><br />Figure 4A. Extracts from HeLa cells containing the indicated amounts of GR (from 30 to 240 fmol), and from 5x10e6 Raji or Molt cells were analysed by Western blot using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). Figure 4B. Western blot analysis of extracts from 300,000 HeLa cells with the Diagenode monoclonal antibody against hGR (concentration 1 μg/ml). </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig5.png" alt="GR Antibody validated in Immunohistochemistry" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig6.png" alt="GR Antibody validated for Immunohistochemistry" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR </strong><br />1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml. <br />2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig7.png" alt="GR Antibody validated in Immunoprecipitation" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR </strong><br />The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above </small></p>
</div>
</div>',
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'price_GBP' => '320',
'price_JPY' => '52900',
'price_CNY' => '',
'price_AUD' => '900',
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'meta_title' => 'GR Antibody - ChIP-seq Grade (C15200010) | Diagenode',
'meta_keywords' => '',
'meta_description' => 'GR (Glucocorticoid receptor) Monoclonal Antibody validated in ChIP-seq, ChIP-qPCR, WB, Flow Cyt, IP and ELISA. Batch-specific data available on the website. Alternative names: NR3C1, GCCR, GCR, GRL. Sample size avaialble.',
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'name' => 'Bioruptor<sup>®</sup> Pico sonication device',
'description' => '<p><a href="https://go.diagenode.com/bioruptor-upgrade"><img src="https://www.diagenode.com/img/banners/banner-br-trade.png" /></a></p>
<p>The Bioruptor® Pico (2013-2019) represented a breakthrough for shearing micro-volumes of 5 μl to larger volumes of up to 2 ml. <span>The new generation keeps the features you like the most and bring even more innovation. Check it now:</span></p>
<center><span></span></center><center><a href="https://www.diagenode.com/p/bioruptorpico2"> <img alt="New Bioruptor Pico" src="https://www.diagenode.com/img/product/shearing_technologies/new-pico-product-banner.jpg" /></a></center>
<p></p>
<p><span>Watch our short video about the Bioruptor Pico and how it can help you accomplish perfect shearing for any application including chromatin shearing, DNA shearing for NGS, unmatched DNA extraction from FFPE samples, RNA shearing, protein extraction, and much more.</span></p>
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'label1' => 'User manual ',
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'label2' => 'Recommended settings for DNA shearing with Bioruptor® Pico',
'info2' => '<p>Follow our guidelines and find the good parameters for your expected DNA size: <a href="https://pybrevet.typeform.com/to/o8cQfM">DNA shearing with the Bioruptor® Pico</a></p>
<p></p>
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'label3' => 'Available chromatin shearing kits',
'info3' => '<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
<table style="width: 925px;">
<tbody>
<tr valign="middle">
<td style="width: 213px;"></td>
<td style="text-align: center; width: 208px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histones)</a></strong></td>
<td style="text-align: center; width: 180px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center; width: 154px;"><strong><a href="../p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center; width: 155px;"><strong><a href="../p/chromatin-shearing-plant-chip-seq-kit">Chromatin Shearing Kit (for Plant)</a></strong></td>
</tr>
<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>SDS concentration</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;">< 0.1%</p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;">0.2%</p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;">1%</p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;">0.5%</p>
</td>
</tr>
<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Nuclei isolation</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;">Yes</p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;">Yes</p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;">No</p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;">Yes</p>
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<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Allows for shearing of... cells/tissue</strong></p>
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<td style="text-align: center; width: 180px;">
<p style="text-align: center;">100 million cells</p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;">100 million cells</p>
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<td style="text-align: center; width: 155px;">
<p style="text-align: center;">up to 25 g of tissue</p>
</td>
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<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
<p style="text-align: center;"><a href="https://www.diagenode.com/en/p/manual-chipmentation-kit-for-histones-24-rxns">ChIPmentation Kit for Histones</a></p>
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<td style="text-align: center; width: 180px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p style="text-align: center;"><a href="../p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;"><a href="../p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;"><a href="../p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant <br />ChIP-seq kit</a></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<div class="small-2 columns"><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></div>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'authors' => 'Nicolaides N. et al.',
'description' => '<p>Background: Many rapid nongenomic glucocorticoid actions are mediated by membrane-bound glucocorticoid receptors (GRs). S-palmitoylation is a lipid post-translational modification that mediates the membrane localization of some steroid receptors. A highly homologous amino acid sequence (663YLCM KTLLL671) is present in the ligand-binding domain of hGRα, suggesting that hGRα might also undergo S-palmitoylation.</p>
<p>Aim: To investigate the role of the motif 663YLCMKTLLL671 in membrane localization of the hGRα and in mediating rapid nongenomic glucocorticoid signaling.</p>
<p>Methods and Results: We showed that the mutant receptors hGRαY663A, hGRαC665A and hGRαLL670/671AA, and the addition of the palmitoylation inhibitor 2-bromopalmitate did not prevent membrane localization of hGRα and co-localization with caveolin-1, and did not influence the biphasic activation of mitogen-activated protein kinase (MAPK) signaling pathway in the early time points. Finally, the hGRα was not shown to undergo S-palmitoylation.</p>
<p>Conclusions: The motif 663YLCMKTLLL671 does not play a role in membrane localization of hGRα and does not mediate the nongenomic glucocorticoid actions.</p>',
'date' => '2017-04-15',
'pmid' => 'http://www.jmolbiochem.com/index.php/JmolBiochem/article/view/199',
'doi' => '',
'modified' => '2017-07-07 16:21:23',
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'name' => 'Three Novel Heterozygous Point Mutations of NR3C1 causing Glucocorticoid Resistance',
'authors' => 'Vitellius G et al.',
'description' => '<p>Generalized glucocorticoid resistance is associated with glucocorticoid receptor (GR, NR3C1) mutations. Three novel heterozygous missense NR3C1 mutations (R477S, Y478C and L672P) were identified in patients presenting with adrenal incidentalomas, glucocorticoid excess without Cushing syndrome. Dexamethasone (DXM) binding studies demonstrated that the affinity of GR<sub>R477S</sub> and GR<sub>Y478C</sub> mutants, located in the DNA-binding domain of GR, was similar to wild-type GR (Kd = 2-3 nM). In contrast, GR<sub>L672P</sub> mutant, located in the ligand-binding domain of GR, was unable to bind glucocorticoids and was more sensitive to protein degradation. GR subcellular distribution revealed a marked decrease in DXM-induced nuclear translocation of GR<sub>R477S</sub> and GR<sub>Y478C</sub> mutants, whereas GR<sub>L672P</sub> remained exclusively cytoplasmic. Chromatin immunoprecipitation demonstrated impaired recruitment of DBD mutants onto the regulatory sequence of FKBP5. Transactivation assays disclosed the lack of transcriptional activity of GR<sub>R477S</sub> and GR<sub>L672P</sub> while GR<sub>Y478C</sub> had a reduced transactivation capacity. Three-D modeling indicated that R477S lost two essential hydrogen bonds with DNA, Y478C resulted in altered interaction with surrounding amino-acids, destabilizing DBD, while L672P altered the H8 helix folding, leading to unstructured LBD. This study identifies novel NR3C1 mutations with their molecular consequences on altered GR signaling and suggests that genetic screening of NR3C1 should be conducted in patients with subclinical hypercorticism.</p>',
'date' => '2016-04-27',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/27120390',
'doi' => '10.1002/humu.23008',
'modified' => '2016-05-13 17:32:26',
'created' => '2016-05-13 17:32:26',
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'id' => '2967',
'name' => 'Functional characterization of the hGRαT556I causing Chrousos syndrome',
'authors' => 'Nicolaides NC et al.',
'description' => '<h4>BACKGROUND:</h4>
<p><abstracttext label="BACKGROUND" nlmcategory="BACKGROUND">Chrousos syndrome is a rare pathologic condition characterized by generalized, partial resistance of target tissues to glucocorticoids and caused by inactivating mutations of the human glucocorticoid receptor (hGR) gene. A novel case of Chrousos syndrome has been reported in a patient with adrenal incidentaloma, who harboured a heterozygous point mutation in the hGR gene, which resulted in threonine (T) to isoleucine (I) substitution at amino acid position 556 in the ligand-binding domain of the receptor.</abstracttext></p>
<h4>OBJECTIVE:</h4>
<p><abstracttext label="OBJECTIVE" nlmcategory="OBJECTIVE">To delineate the molecular mechanisms through which the mutant receptor hGRαT556I causes Chrousos syndrome.</abstracttext></p>
<h4>DESIGN AND RESULTS:</h4>
<p><abstracttext label="DESIGN AND RESULTS" nlmcategory="RESULTS">Compared with the wild-type receptor, the mutant receptor hGRαT556I demonstrated 50% reduction in its ability to transactivate glucocorticoid-responsive genes and in the affinity for the ligand, 30% increase in the ability to transrepress the nuclear factor-κB-target genes and a 3,4-fold delay in the cytoplasmic-to-nuclear translocation. The mutant receptor hGRαT556I did not exert a dominant negative effect upon the hGRα-mediated transcriptional activity; it preserved its ability to bind to DNA and interacted with the glucocorticoid receptor-interacting protein 1 coactivator mostly through its activation function-1 domain. Structural biology studies revealed that the T556I mutation caused disruption of the hydrogen bond formed by the T556 with the =O group of P637 backbone, which resulted in a significant relocation of the P637-bearing loop. This conformational alteration affected the local 3D arrangement of the receptor and hence the electrostatic surface of the region.</abstracttext></p>
<h4>CONCLUSIONS:</h4>
<p><abstracttext label="CONCLUSIONS" nlmcategory="CONCLUSIONS">The hGRαT556I causes Chrousos syndrome by impairing multiple steps of the glucocorticoid signal transduction pathway.</abstracttext></p>',
'date' => '2016-01-01',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/26541474',
'doi' => ' 10.1111/eci.12563',
'modified' => '2016-06-29 09:41:42',
'created' => '2016-06-29 09:41:42',
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'id' => '2939',
'name' => 'Transient generalized glucocorticoid hypersensitivity',
'authors' => 'Nicolaides NC et al.',
'description' => '<h4>BACKGROUND:</h4>
<p><abstracttext label="BACKGROUND" nlmcategory="BACKGROUND">Transient generalized glucocorticoid hypersensitivity is a rare disorder characterized by increased tissue sensitivity to glucocorticoids and compensatory hypo-activation of the hypothalamic-pituitary-adrenal axis. The condition itself and the underlying molecular mechanisms have not been elucidated.</abstracttext></p>
<h4>OBJECTIVE:</h4>
<p><abstracttext label="OBJECTIVE" nlmcategory="OBJECTIVE">To present the clinical manifestations, endocrinologic evaluation and transcriptomic profile in a patient with transient generalized glucocorticoid hypersensitivity.</abstracttext></p>
<h4>DESIGN AND RESULTS:</h4>
<p><abstracttext label="DESIGN AND RESULTS" nlmcategory="RESULTS">A 9-year-old girl presented with an 8-month history of clinical manifestations suggestive of Cushing syndrome. Endocrinologic evaluation revealed undetectable 08:00 h ACTH (<1 pg/mL) and cortisol (0·025 μg/dL) concentrations, which remained decreased throughout the 24-h period and did not respond to stimulation with ovine CRH. The disease gradually resolved spontaneously over the ensuing 3 months. Sequencing of the human glucocorticoid receptor gene revealed no mutations or polymorphisms. Western blot analysis in peripheral blood mononuclear cells revealed equal protein expression of hGRα of the patient in the disease and postresolution phases compared with a control subject. Transcriptomic analysis in peripheral blood mononuclear cells in the disease and postresolution phases identified 903 differentially expressed genes. Of these, 106 genes were up-regulated and 797 were down-regulated in the disease compared with the resolution phase. Bioinformatics analysis on the differentially expressed gene networks revealed Nuclear Factor-κB as the predominant transcription factor influencing the expression of the majority of differentially expressed genes.</abstracttext></p>
<h4>CONCLUSIONS:</h4>
<p><abstracttext label="CONCLUSIONS" nlmcategory="CONCLUSIONS">Our findings indicate that a transient postreceptor defect, or a virus- or bacterium-encoded molecule, may have enhanced glucocorticoid signal transduction, leading to transient generalized glucocorticoid hypersensitivity and hypo-activation of the HPA axis.</abstracttext></p>',
'date' => '2015-12-01',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/26479047',
'doi' => ' 10.1111/eci.12554',
'modified' => '2016-05-27 10:41:26',
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'name' => 'Perinatal exposure to low-dose bisphenol A affects the neuroendocrine stress response in rats.',
'authors' => 'Panagiotidou E, Zerva S, Mitsiou DJ, Alexis MN, Kitraki E',
'description' => 'Bisphenol A (BPA) is an estrogen-mimicking endocrine disruptor. Early-life exposures to low doses of BPA exert long-lasting effects on animals' reproductive and brain physiology. However, little is known about the effects of BPA on the stress-response system. Given the interaction of sex and stress hormones, we examined the effect of a low perinatal BPA exposure on the function of the hypothalamic-pituitary-adrenal (HPA) axis at rest and upon application of acute stress. Throughout pregnancy and lactation rats received daily 40 μg BPA/kg body weight orally via cornflakes. We studied the effect of this low but chronic exposure to BPA in the male and female offspring at puberty. BPA exposure led to abnormal adrenal histology including reduced zona reticularis especially in male offspring, hyperplasia of zona fasciculata in both sexes, and increased adrenal weight in female offspring. BPA-treated females had increased basal corticosterone and reduced hypothalamic glucocorticoid receptors (GR) levels. Stressed BPA-exposed females exhibited anxiety-like behavioral coping, a less rigorous corticosterone response, and did not downregulate GR in the hypothalamus, compared with control females. BPA-exposed males exhibited a heightened corticosterone stress response compared with females; they also displayed increased pro-opiomelanocortin mRNA levels and retained the prestress levels of pituitary corticotropin-releasing hormone-receptor 1, compared with control males. We found that perinatal chronic exposure to a low dose of BPA perturbs the basal and stress-induced activity of the HPA axis in a sexually dimorphic manner at adolescence. Exposure to BPA might contribute to increased susceptibility to stress-related disorders in later life.',
'date' => '2014-03-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/24323913',
'doi' => '',
'modified' => '2015-07-24 15:39:01',
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'name' => 'Pattern of heat shock factor and heat shock protein expression in lymphocytes of bipolar patients: Increased HSP70-glucocorticoid receptor heterocomplex.',
'authors' => 'Bei ES, Salpeas V, Alevizos B, Anagnostara C, Pappa D, Moutsatsou P',
'description' => 'Bipolar disorder (BD), a stress-related disease, is characterized by altered glucocorticoid receptor (GR) signalling. Stress response includes activation of heat shock factor (HSF) and subsequent heat shock protein (HSP) synthesis which regulate GR folding and function. The objective of this study was to investigate the possible role of HSFs, HSPs and their interaction with GR in BD. We applied immunoprecipitation, SDS-PAGE/Western blot analysis and electrophoretic mobility shift assay (EMSA) in lymphocytes (whole cell or nuclear extracts) from BD patients and healthy subjects and determined the HSPs (HSP90 and HSP70), the heterocomplexes HSP90-GR and HSP70-GR, the HSFs (HSF1 and HSF4) as well as the HSF-DNA binding. The HSP70-GR heterocomplex was elevated (p < 0.05) in BD patients vs healthy subjects, and nuclear HSP70 was reduced (p ≤ 0.01) in bipolar manic patients. Protein levels of HSF1, HSF4, HSP90, HSP90-GR heterocomplex, and HSF-DNA binding remained unaltered in BD patients vs healthy subjects. The corresponding effect sizes (ES) indicated a large ES for HSP70-GR, HSP70, HSF-DNA binding and HSF4, and a medium ES for HSP90, HSF1 and HSP90-GR between healthy subjects and bipolar patients. Significant correlations among HSFs, HSPs, GR and HSP70-GR heterocomplex were observed in healthy subjects, which were abrogated in bipolar patients. The higher interaction between GR and HSP70 and the disturbances in the relations among heat shock response parameters and GR as observed in our BD patients may provide novel insights into the contribution of these factors in BD aetiopathogenesis.',
'date' => '2013-11-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/23938235',
'doi' => '',
'modified' => '2015-07-24 15:39:00',
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'name' => 'Coactivation of GR and NFKB alters the repertoire of their binding sites and target genes.',
'authors' => 'Rao NA, McCalman MT, Moulos P, Francoijs KJ, Chatziioannou A, Kolisis FN, Alexis MN, Mitsiou DJ, Stunnenberg HG',
'description' => 'Glucocorticoid receptor (GR) exerts anti-inflammatory action in part by antagonizing proinflammatory transcription factors such as the nuclear factor kappa-b (NFKB). Here, we assess the crosstalk of activated GR and RELA (p65, major NFKB component) by global identification of their binding sites and target genes. We show that coactivation of GR and p65 alters the repertoire of regulated genes and results in their association with novel sites in a mutually dependent manner. These novel sites predominantly cluster with p65 target genes that are antagonized by activated GR and vice versa. Our data show that coactivation of GR and NFKB alters signaling pathways that are regulated by each factor separately and provide insight into the networks underlying the GR and NFKB crosstalk.',
'date' => '2011-09-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/21750107',
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<p><span>Watch our short video about the Bioruptor Pico and how it can help you accomplish perfect shearing for any application including chromatin shearing, DNA shearing for NGS, unmatched DNA extraction from FFPE samples, RNA shearing, protein extraction, and much more.</span></p>
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'info3' => '<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
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<tbody>
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<td style="width: 213px;"></td>
<td style="text-align: center; width: 208px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histones)</a></strong></td>
<td style="text-align: center; width: 180px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center; width: 154px;"><strong><a href="../p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center; width: 155px;"><strong><a href="../p/chromatin-shearing-plant-chip-seq-kit">Chromatin Shearing Kit (for Plant)</a></strong></td>
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<p style="text-align: center;">< 0.1%</p>
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<p style="text-align: center;">0.2%</p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;">1%</p>
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<td style="text-align: center; width: 155px;">
<p style="text-align: center;">0.5%</p>
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<td style="text-align: center; width: 180px;">
<p style="text-align: center;">Yes</p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;">No</p>
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<td style="text-align: center; width: 155px;">
<p style="text-align: center;">Yes</p>
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<p style="text-align: center;">100 million cells</p>
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<p style="text-align: center;">100 million cells</p>
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<p style="text-align: center;">100 million cells</p>
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<p style="text-align: center;">up to 25 g of tissue</p>
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<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
<p style="text-align: center;"><a href="https://www.diagenode.com/en/p/manual-chipmentation-kit-for-histones-24-rxns">ChIPmentation Kit for Histones</a></p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p style="text-align: center;"><a href="../p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;"><a href="../p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;"><a href="../p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant <br />ChIP-seq kit</a></p>
</td>
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<p>Monoclonal antibody raised in mouse against amino acids 304-428 of the human <strong>GR (glucocorticoid receptor),</strong> using a chimeric protein.</p>',
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'description' => 'Glucocorticoid receptor (GR) exerts anti-inflammatory action in part by antagonizing proinflammatory transcription factors such as the nuclear factor kappa-b (NFKB). Here, we assess the crosstalk of activated GR and RELA (p65, major NFKB component) by global identification of their binding sites and target genes. We show that coactivation of GR and p65 alters the repertoire of regulated genes and results in their association with novel sites in a mutually dependent manner. These novel sites predominantly cluster with p65 target genes that are antagonized by activated GR and vice versa. Our data show that coactivation of GR and NFKB alters signaling pathways that are regulated by each factor separately and provide insight into the networks underlying the GR and NFKB crosstalk.',
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP assays were performed using HeLa cells, the Diagenode monoclonal antibody directed against GR (Cat. No. MAb-010-050) and optimized PCR primer sets for qPCR. The cells were treated either with ethanol (EtOH, used as a negative control) or triamcinolone acetonide (TA) for 4 hours prior to cell harvesting. ChIP was performed using sheared chromatin from 3 million cells and 5 μg of antibody. QPCR was performed with primers for the human metallothionein promoter (hMTIIA) and for exon 2 of the human myoglobin gene (hmyo ex2), used as a negative control. Figure 1 shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +/- control target). These results demonstrate the occupancy of the human metallothionein IIA promoter by GR. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2A.png" alt="GR Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2B.png" alt="GR Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2C.png" alt="GR Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2D.png" alt="GR Antibody validated in ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig2E.png" alt="GR Antibody ChIP-seq Grade" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP was performed on sheared chromatin from 3.5 million HeLaB2 cells using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). The cells were treated with the synthetic GR ligand triamcinolone acetonide (TA) for 4 hours prior to harvesting. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence of chromosome 16 (figure 2A) as well as the MT2A positive control gene (figure 2B). The position of the PCR amplicon is also indicated. Figure 2C, D and E show the results for the known GR target genes PER1 on chromosome 17 and FKBP5 and TNFAIP3 on chromosome 6. </small></p>
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<p><small><strong> Figure 3. Sandwich ELISA </strong><br />The specificity of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050) was assessed by sandwich ELISA. Figure 3A: schematic representation of the sandwich ELISA with the monoclonal antibody against hGR (clone #:m2F8). Figure 3B: ELISA results using the monoclonal antibody against hGR at a concentration of 0.5 μg/ml. The figure shows an ELISA signal which is proportionally increasing with increasing amounts of recombinant hGR. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody against hGR </strong><br />Figure 4A. Extracts from HeLa cells containing the indicated amounts of GR (from 30 to 240 fmol), and from 5x10e6 Raji or Molt cells were analysed by Western blot using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). Figure 4B. Western blot analysis of extracts from 300,000 HeLa cells with the Diagenode monoclonal antibody against hGR (concentration 1 μg/ml). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig6.png" alt="GR Antibody validated for Immunohistochemistry" /></p>
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<p><small><strong> Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR </strong><br />1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml. <br />2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml. </small></p>
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<p><small><strong> Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR </strong><br />The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP assays were performed using HeLa cells, the Diagenode monoclonal antibody directed against GR (Cat. No. MAb-010-050) and optimized PCR primer sets for qPCR. The cells were treated either with ethanol (EtOH, used as a negative control) or triamcinolone acetonide (TA) for 4 hours prior to cell harvesting. ChIP was performed using sheared chromatin from 3 million cells and 5 μg of antibody. QPCR was performed with primers for the human metallothionein promoter (hMTIIA) and for exon 2 of the human myoglobin gene (hmyo ex2), used as a negative control. Figure 1 shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +/- control target). These results demonstrate the occupancy of the human metallothionein IIA promoter by GR. </small></p>
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<p><small><strong> Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR </strong><br />1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml. <br />2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml. </small></p>
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<p><small><strong> Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR </strong><br />The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against hGR </strong><br />ChIP was performed on sheared chromatin from 3.5 million HeLaB2 cells using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). The cells were treated with the synthetic GR ligand triamcinolone acetonide (TA) for 4 hours prior to harvesting. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence of chromosome 16 (figure 2A) as well as the MT2A positive control gene (figure 2B). The position of the PCR amplicon is also indicated. Figure 2C, D and E show the results for the known GR target genes PER1 on chromosome 17 and FKBP5 and TNFAIP3 on chromosome 6. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig3.png" alt="GR Antibody ELISA Validation" /></p>
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<div class="small-6 columns">
<p><small><strong> Figure 3. Sandwich ELISA </strong><br />The specificity of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050) was assessed by sandwich ELISA. Figure 3A: schematic representation of the sandwich ELISA with the monoclonal antibody against hGR (clone #:m2F8). Figure 3B: ELISA results using the monoclonal antibody against hGR at a concentration of 0.5 μg/ml. The figure shows an ELISA signal which is proportionally increasing with increasing amounts of recombinant hGR. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig4.png" alt="GR Antibody validated in Western Blot" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody against hGR </strong><br />Figure 4A. Extracts from HeLa cells containing the indicated amounts of GR (from 30 to 240 fmol), and from 5x10e6 Raji or Molt cells were analysed by Western blot using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). Figure 4B. Western blot analysis of extracts from 300,000 HeLa cells with the Diagenode monoclonal antibody against hGR (concentration 1 μg/ml). </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig5.png" alt="GR Antibody validated in Immunohistochemistry" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig6.png" alt="GR Antibody validated for Immunohistochemistry" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR </strong><br />1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml. <br />2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml. </small></p>
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</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200010_fig7.png" alt="GR Antibody validated in Immunoprecipitation" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR </strong><br />The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above </small></p>
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'meta_description' => 'GR (Glucocorticoid receptor) Monoclonal Antibody validated in ChIP-seq, ChIP-qPCR, WB, Flow Cyt, IP and ELISA. Batch-specific data available on the website. Alternative names: NR3C1, GCCR, GCR, GRL. Sample size avaialble.',
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'description' => '<p><a href="https://go.diagenode.com/bioruptor-upgrade"><img src="https://www.diagenode.com/img/banners/banner-br-trade.png" /></a></p>
<p>The Bioruptor® Pico (2013-2019) represented a breakthrough for shearing micro-volumes of 5 μl to larger volumes of up to 2 ml. <span>The new generation keeps the features you like the most and bring even more innovation. Check it now:</span></p>
<center><span></span></center><center><a href="https://www.diagenode.com/p/bioruptorpico2"> <img alt="New Bioruptor Pico" src="https://www.diagenode.com/img/product/shearing_technologies/new-pico-product-banner.jpg" /></a></center>
<p></p>
<p><span>Watch our short video about the Bioruptor Pico and how it can help you accomplish perfect shearing for any application including chromatin shearing, DNA shearing for NGS, unmatched DNA extraction from FFPE samples, RNA shearing, protein extraction, and much more.</span></p>
<p>
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<p></p>
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'label3' => 'Available chromatin shearing kits',
'info3' => '<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
<table style="width: 925px;">
<tbody>
<tr valign="middle">
<td style="width: 213px;"></td>
<td style="text-align: center; width: 208px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histones)</a></strong></td>
<td style="text-align: center; width: 180px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center; width: 154px;"><strong><a href="../p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center; width: 155px;"><strong><a href="../p/chromatin-shearing-plant-chip-seq-kit">Chromatin Shearing Kit (for Plant)</a></strong></td>
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<td style="width: 213px;">
<p style="text-align: left;"><strong>SDS concentration</strong></p>
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<p style="text-align: center;">< 0.1%</p>
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<td style="text-align: center; width: 180px;">
<p style="text-align: center;">0.2%</p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;">1%</p>
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<td style="text-align: center; width: 155px;">
<p style="text-align: center;">0.5%</p>
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<td style="width: 213px;">
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<p style="text-align: center;">Yes</p>
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<td style="text-align: center; width: 180px;">
<p style="text-align: center;">Yes</p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;">No</p>
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<td style="text-align: center; width: 155px;">
<p style="text-align: center;">Yes</p>
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<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
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<td style="text-align: center; width: 208px;">
<p style="text-align: center;">100 million cells</p>
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<td style="text-align: center; width: 180px;">
<p style="text-align: center;">100 million cells</p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;">100 million cells</p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;">up to 25 g of tissue</p>
</td>
</tr>
<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
<p style="text-align: center;"><a href="https://www.diagenode.com/en/p/manual-chipmentation-kit-for-histones-24-rxns">ChIPmentation Kit for Histones</a></p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p style="text-align: center;"><a href="../p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;"><a href="../p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;"><a href="../p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant <br />ChIP-seq kit</a></p>
</td>
</tr>
</tbody>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
<div class="small-12 medium-3 large-3 columns">
<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
</div>
</div>
<p>Diagenode’s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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'description' => '<p><span style="font-weight: 400;">All Diagenode’s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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'meta_title' => 'Diagenode's selection of Antibodies is exclusively dedicated for Epigenetic Research | Diagenode',
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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'name' => 'ChIP-grade antibodies',
'description' => '<div class="row">
<div class="small-10 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
</div>
<div class="small-2 columns"><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></div>
</div>
<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
<div class="row">
<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
<div class="small-12 medium-6 large-6 columns">
<p></p>
<p></p>
<p></p>
</div>
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<p></p>
<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'meta_description' => 'Diagenode Offers Extensively Validated ChIP-Grade Antibodies, Confirmed for their Specificity, and high level of Performance in Chromatin Immunoprecipitation ChIP',
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'url' => 'files/products/antibodies/Datasheet_hGR_C15200010.pdf',
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'name' => 'Antibodies you can trust',
'description' => '<p style="text-align: justify;"><span>Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of large sets of genome-wide sequencing data. ChIP sequencing (ChIP-seq) has now become the gold standard method for chromatin studies, given the accuracy and coverage scale of the approach over other methods. Successful ChIP-seq, however, requires a higher level of experimental accuracy and consistency in all steps of ChIP than ever before. Particularly crucial is the quality of ChIP antibodies. </span></p>',
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'id' => '38',
'name' => 'Epigenetic Antibodies Brochure',
'description' => '<p>More than in any other immuoprecipitation assays, quality antibodies are critical tools in many epigenetics experiments. Since 10 years, Diagenode has developed the most stringent quality production available on the market for antibodies exclusively focused on epigenetic uses. All our antibodies have been qualified to work in epigenetic applications.</p>',
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'name' => 'Coordinated glucocorticoid receptor and MAFB action inducestolerogenesis and epigenome remodeling in dendritic cells',
'authors' => 'Morante-Palacios Octavio et al.',
'description' => '<p>Abstract Glucocorticoids (GCs) exert potent anti-inflammatory effects in immune cells through the glucocorticoid receptor (GR). Dendritic cells (DCs), central actors for coordinating immune responses, acquire tolerogenic properties in response to GCs. Tolerogenic DCs (tolDCs) have emerged as a potential treatment for various inflammatory diseases. To date, the underlying cell type-specific regulatory mechanisms orchestrating GC-mediated acquisition of immunosuppressive properties remain poorly understood. In this study, we investigated the transcriptomic and epigenomic remodeling associated with differentiation to DCs in the presence of GCs. Our analysis demonstrates a major role of MAFB in this process, in synergy with GR. GR and MAFB both interact with methylcytosine dioxygenase TET2 and bind to genomic loci that undergo specific demethylation in tolDCs. We also show that the role of MAFB is more extensive, binding to thousands of genomic loci in tolDCs. Finally, MAFB knockdown erases the tolerogenic properties of tolDCs and reverts the specific DNA demethylation and gene upregulation. The preeminent role of MAFB is also demonstrated in vivo for myeloid cells from synovium in rheumatoid arthritis following GC treatment. Our results imply that, once directly activated by GR, MAFB plays a critical role in orchestrating the epigenomic and transcriptomic remodeling that define the tolerogenic phenotype.</p>',
'date' => '2022-01-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/34893889',
'doi' => '10.1093/nar/gkab1182',
'modified' => '2022-05-20 09:44:29',
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'id' => '3208',
'name' => 'The Role of S-Palmitoylation of the Human Glucocorticoid Receptor (hGR) in Mediating the Nongenomic Glucocorticoid Actions',
'authors' => 'Nicolaides N. et al.',
'description' => '<p>Background: Many rapid nongenomic glucocorticoid actions are mediated by membrane-bound glucocorticoid receptors (GRs). S-palmitoylation is a lipid post-translational modification that mediates the membrane localization of some steroid receptors. A highly homologous amino acid sequence (663YLCM KTLLL671) is present in the ligand-binding domain of hGRα, suggesting that hGRα might also undergo S-palmitoylation.</p>
<p>Aim: To investigate the role of the motif 663YLCMKTLLL671 in membrane localization of the hGRα and in mediating rapid nongenomic glucocorticoid signaling.</p>
<p>Methods and Results: We showed that the mutant receptors hGRαY663A, hGRαC665A and hGRαLL670/671AA, and the addition of the palmitoylation inhibitor 2-bromopalmitate did not prevent membrane localization of hGRα and co-localization with caveolin-1, and did not influence the biphasic activation of mitogen-activated protein kinase (MAPK) signaling pathway in the early time points. Finally, the hGRα was not shown to undergo S-palmitoylation.</p>
<p>Conclusions: The motif 663YLCMKTLLL671 does not play a role in membrane localization of hGRα and does not mediate the nongenomic glucocorticoid actions.</p>',
'date' => '2017-04-15',
'pmid' => 'http://www.jmolbiochem.com/index.php/JmolBiochem/article/view/199',
'doi' => '',
'modified' => '2017-07-07 16:21:23',
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(int) 2 => array(
'id' => '2923',
'name' => 'Three Novel Heterozygous Point Mutations of NR3C1 causing Glucocorticoid Resistance',
'authors' => 'Vitellius G et al.',
'description' => '<p>Generalized glucocorticoid resistance is associated with glucocorticoid receptor (GR, NR3C1) mutations. Three novel heterozygous missense NR3C1 mutations (R477S, Y478C and L672P) were identified in patients presenting with adrenal incidentalomas, glucocorticoid excess without Cushing syndrome. Dexamethasone (DXM) binding studies demonstrated that the affinity of GR<sub>R477S</sub> and GR<sub>Y478C</sub> mutants, located in the DNA-binding domain of GR, was similar to wild-type GR (Kd = 2-3 nM). In contrast, GR<sub>L672P</sub> mutant, located in the ligand-binding domain of GR, was unable to bind glucocorticoids and was more sensitive to protein degradation. GR subcellular distribution revealed a marked decrease in DXM-induced nuclear translocation of GR<sub>R477S</sub> and GR<sub>Y478C</sub> mutants, whereas GR<sub>L672P</sub> remained exclusively cytoplasmic. Chromatin immunoprecipitation demonstrated impaired recruitment of DBD mutants onto the regulatory sequence of FKBP5. Transactivation assays disclosed the lack of transcriptional activity of GR<sub>R477S</sub> and GR<sub>L672P</sub> while GR<sub>Y478C</sub> had a reduced transactivation capacity. Three-D modeling indicated that R477S lost two essential hydrogen bonds with DNA, Y478C resulted in altered interaction with surrounding amino-acids, destabilizing DBD, while L672P altered the H8 helix folding, leading to unstructured LBD. This study identifies novel NR3C1 mutations with their molecular consequences on altered GR signaling and suggests that genetic screening of NR3C1 should be conducted in patients with subclinical hypercorticism.</p>',
'date' => '2016-04-27',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/27120390',
'doi' => '10.1002/humu.23008',
'modified' => '2016-05-13 17:32:26',
'created' => '2016-05-13 17:32:26',
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'id' => '2967',
'name' => 'Functional characterization of the hGRαT556I causing Chrousos syndrome',
'authors' => 'Nicolaides NC et al.',
'description' => '<h4>BACKGROUND:</h4>
<p><abstracttext label="BACKGROUND" nlmcategory="BACKGROUND">Chrousos syndrome is a rare pathologic condition characterized by generalized, partial resistance of target tissues to glucocorticoids and caused by inactivating mutations of the human glucocorticoid receptor (hGR) gene. A novel case of Chrousos syndrome has been reported in a patient with adrenal incidentaloma, who harboured a heterozygous point mutation in the hGR gene, which resulted in threonine (T) to isoleucine (I) substitution at amino acid position 556 in the ligand-binding domain of the receptor.</abstracttext></p>
<h4>OBJECTIVE:</h4>
<p><abstracttext label="OBJECTIVE" nlmcategory="OBJECTIVE">To delineate the molecular mechanisms through which the mutant receptor hGRαT556I causes Chrousos syndrome.</abstracttext></p>
<h4>DESIGN AND RESULTS:</h4>
<p><abstracttext label="DESIGN AND RESULTS" nlmcategory="RESULTS">Compared with the wild-type receptor, the mutant receptor hGRαT556I demonstrated 50% reduction in its ability to transactivate glucocorticoid-responsive genes and in the affinity for the ligand, 30% increase in the ability to transrepress the nuclear factor-κB-target genes and a 3,4-fold delay in the cytoplasmic-to-nuclear translocation. The mutant receptor hGRαT556I did not exert a dominant negative effect upon the hGRα-mediated transcriptional activity; it preserved its ability to bind to DNA and interacted with the glucocorticoid receptor-interacting protein 1 coactivator mostly through its activation function-1 domain. Structural biology studies revealed that the T556I mutation caused disruption of the hydrogen bond formed by the T556 with the =O group of P637 backbone, which resulted in a significant relocation of the P637-bearing loop. This conformational alteration affected the local 3D arrangement of the receptor and hence the electrostatic surface of the region.</abstracttext></p>
<h4>CONCLUSIONS:</h4>
<p><abstracttext label="CONCLUSIONS" nlmcategory="CONCLUSIONS">The hGRαT556I causes Chrousos syndrome by impairing multiple steps of the glucocorticoid signal transduction pathway.</abstracttext></p>',
'date' => '2016-01-01',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/26541474',
'doi' => ' 10.1111/eci.12563',
'modified' => '2016-06-29 09:41:42',
'created' => '2016-06-29 09:41:42',
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(int) 4 => array(
'id' => '2939',
'name' => 'Transient generalized glucocorticoid hypersensitivity',
'authors' => 'Nicolaides NC et al.',
'description' => '<h4>BACKGROUND:</h4>
<p><abstracttext label="BACKGROUND" nlmcategory="BACKGROUND">Transient generalized glucocorticoid hypersensitivity is a rare disorder characterized by increased tissue sensitivity to glucocorticoids and compensatory hypo-activation of the hypothalamic-pituitary-adrenal axis. The condition itself and the underlying molecular mechanisms have not been elucidated.</abstracttext></p>
<h4>OBJECTIVE:</h4>
<p><abstracttext label="OBJECTIVE" nlmcategory="OBJECTIVE">To present the clinical manifestations, endocrinologic evaluation and transcriptomic profile in a patient with transient generalized glucocorticoid hypersensitivity.</abstracttext></p>
<h4>DESIGN AND RESULTS:</h4>
<p><abstracttext label="DESIGN AND RESULTS" nlmcategory="RESULTS">A 9-year-old girl presented with an 8-month history of clinical manifestations suggestive of Cushing syndrome. Endocrinologic evaluation revealed undetectable 08:00 h ACTH (<1 pg/mL) and cortisol (0·025 μg/dL) concentrations, which remained decreased throughout the 24-h period and did not respond to stimulation with ovine CRH. The disease gradually resolved spontaneously over the ensuing 3 months. Sequencing of the human glucocorticoid receptor gene revealed no mutations or polymorphisms. Western blot analysis in peripheral blood mononuclear cells revealed equal protein expression of hGRα of the patient in the disease and postresolution phases compared with a control subject. Transcriptomic analysis in peripheral blood mononuclear cells in the disease and postresolution phases identified 903 differentially expressed genes. Of these, 106 genes were up-regulated and 797 were down-regulated in the disease compared with the resolution phase. Bioinformatics analysis on the differentially expressed gene networks revealed Nuclear Factor-κB as the predominant transcription factor influencing the expression of the majority of differentially expressed genes.</abstracttext></p>
<h4>CONCLUSIONS:</h4>
<p><abstracttext label="CONCLUSIONS" nlmcategory="CONCLUSIONS">Our findings indicate that a transient postreceptor defect, or a virus- or bacterium-encoded molecule, may have enhanced glucocorticoid signal transduction, leading to transient generalized glucocorticoid hypersensitivity and hypo-activation of the HPA axis.</abstracttext></p>',
'date' => '2015-12-01',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/26479047',
'doi' => ' 10.1111/eci.12554',
'modified' => '2016-05-27 10:41:26',
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'name' => 'Perinatal exposure to low-dose bisphenol A affects the neuroendocrine stress response in rats.',
'authors' => 'Panagiotidou E, Zerva S, Mitsiou DJ, Alexis MN, Kitraki E',
'description' => 'Bisphenol A (BPA) is an estrogen-mimicking endocrine disruptor. Early-life exposures to low doses of BPA exert long-lasting effects on animals' reproductive and brain physiology. However, little is known about the effects of BPA on the stress-response system. Given the interaction of sex and stress hormones, we examined the effect of a low perinatal BPA exposure on the function of the hypothalamic-pituitary-adrenal (HPA) axis at rest and upon application of acute stress. Throughout pregnancy and lactation rats received daily 40 μg BPA/kg body weight orally via cornflakes. We studied the effect of this low but chronic exposure to BPA in the male and female offspring at puberty. BPA exposure led to abnormal adrenal histology including reduced zona reticularis especially in male offspring, hyperplasia of zona fasciculata in both sexes, and increased adrenal weight in female offspring. BPA-treated females had increased basal corticosterone and reduced hypothalamic glucocorticoid receptors (GR) levels. Stressed BPA-exposed females exhibited anxiety-like behavioral coping, a less rigorous corticosterone response, and did not downregulate GR in the hypothalamus, compared with control females. BPA-exposed males exhibited a heightened corticosterone stress response compared with females; they also displayed increased pro-opiomelanocortin mRNA levels and retained the prestress levels of pituitary corticotropin-releasing hormone-receptor 1, compared with control males. We found that perinatal chronic exposure to a low dose of BPA perturbs the basal and stress-induced activity of the HPA axis in a sexually dimorphic manner at adolescence. Exposure to BPA might contribute to increased susceptibility to stress-related disorders in later life.',
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'description' => 'Bipolar disorder (BD), a stress-related disease, is characterized by altered glucocorticoid receptor (GR) signalling. Stress response includes activation of heat shock factor (HSF) and subsequent heat shock protein (HSP) synthesis which regulate GR folding and function. The objective of this study was to investigate the possible role of HSFs, HSPs and their interaction with GR in BD. We applied immunoprecipitation, SDS-PAGE/Western blot analysis and electrophoretic mobility shift assay (EMSA) in lymphocytes (whole cell or nuclear extracts) from BD patients and healthy subjects and determined the HSPs (HSP90 and HSP70), the heterocomplexes HSP90-GR and HSP70-GR, the HSFs (HSF1 and HSF4) as well as the HSF-DNA binding. The HSP70-GR heterocomplex was elevated (p < 0.05) in BD patients vs healthy subjects, and nuclear HSP70 was reduced (p ≤ 0.01) in bipolar manic patients. Protein levels of HSF1, HSF4, HSP90, HSP90-GR heterocomplex, and HSF-DNA binding remained unaltered in BD patients vs healthy subjects. The corresponding effect sizes (ES) indicated a large ES for HSP70-GR, HSP70, HSF-DNA binding and HSF4, and a medium ES for HSP90, HSF1 and HSP90-GR between healthy subjects and bipolar patients. Significant correlations among HSFs, HSPs, GR and HSP70-GR heterocomplex were observed in healthy subjects, which were abrogated in bipolar patients. The higher interaction between GR and HSP70 and the disturbances in the relations among heat shock response parameters and GR as observed in our BD patients may provide novel insights into the contribution of these factors in BD aetiopathogenesis.',
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'description' => 'Glucocorticoid receptor (GR) exerts anti-inflammatory action in part by antagonizing proinflammatory transcription factors such as the nuclear factor kappa-b (NFKB). Here, we assess the crosstalk of activated GR and RELA (p65, major NFKB component) by global identification of their binding sites and target genes. We show that coactivation of GR and p65 alters the repertoire of regulated genes and results in their association with novel sites in a mutually dependent manner. These novel sites predominantly cluster with p65 target genes that are antagonized by activated GR and vice versa. Our data show that coactivation of GR and NFKB alters signaling pathways that are regulated by each factor separately and provide insight into the networks underlying the GR and NFKB crosstalk.',
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<p>The Bioruptor® Pico (2013-2019) represented a breakthrough for shearing micro-volumes of 5 μl to larger volumes of up to 2 ml. <span>The new generation keeps the features you like the most and bring even more innovation. Check it now:</span></p>
<center><span></span></center><center><a href="https://www.diagenode.com/p/bioruptorpico2"> <img alt="New Bioruptor Pico" src="https://www.diagenode.com/img/product/shearing_technologies/new-pico-product-banner.jpg" /></a></center>
<p></p>
<p><span>Watch our short video about the Bioruptor Pico and how it can help you accomplish perfect shearing for any application including chromatin shearing, DNA shearing for NGS, unmatched DNA extraction from FFPE samples, RNA shearing, protein extraction, and much more.</span></p>
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'label2' => 'Recommended settings for DNA shearing with Bioruptor® Pico',
'info2' => '<p>Follow our guidelines and find the good parameters for your expected DNA size: <a href="https://pybrevet.typeform.com/to/o8cQfM">DNA shearing with the Bioruptor® Pico</a></p>
<p></p>
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'label3' => 'Available chromatin shearing kits',
'info3' => '<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
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<td style="text-align: center; width: 208px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histones)</a></strong></td>
<td style="text-align: center; width: 180px;"><strong><a href="../p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center; width: 154px;"><strong><a href="../p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center; width: 155px;"><strong><a href="../p/chromatin-shearing-plant-chip-seq-kit">Chromatin Shearing Kit (for Plant)</a></strong></td>
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<p style="text-align: center;">< 0.1%</p>
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<p style="text-align: center;">0.2%</p>
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<td style="text-align: center; width: 154px;">
<p style="text-align: center;">1%</p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;">0.5%</p>
</td>
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<tr style="background-color: #fff;" valign="middle">
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<p style="text-align: left;"><strong>Nuclei isolation</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;">Yes</p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;">Yes</p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;">No</p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;">Yes</p>
</td>
</tr>
<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Allows for shearing of... cells/tissue</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;">100 million cells</p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;">100 million cells</p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;">100 million cells</p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;">up to 25 g of tissue</p>
</td>
</tr>
<tr style="background-color: #fff;" valign="middle">
<td style="width: 213px;">
<p style="text-align: left;"><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center; width: 208px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
<p style="text-align: center;"><a href="https://www.diagenode.com/en/p/manual-chipmentation-kit-for-histones-24-rxns">ChIPmentation Kit for Histones</a></p>
</td>
<td style="text-align: center; width: 180px;">
<p style="text-align: center;"><a href="../p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p style="text-align: center;"><a href="../p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center; width: 154px;">
<p style="text-align: center;"><a href="../p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center; width: 155px;">
<p style="text-align: center;"><a href="../p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant <br />ChIP-seq kit</a></p>
</td>
</tr>
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