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<p><strong>pA-Tn5 Transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For flexibility, the fusion protein is not pre-loaded with sequencing adapters. Please follow the recommended <a href="https://www.diagenode.com/en/documents/assembly-protocol-patn5-20042020-rms" target="_blank">assembly protocol</a> prior to use in CUT&Tag or similar assays (ACT-seq, CoBATCH, TIP-seq and other).</p>
<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
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<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
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<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 Transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For flexibility, the fusion protein is not pre-loaded with sequencing adapters. Please follow the recommended <a href="https://www.diagenode.com/en/documents/assembly-protocol-patn5-20042020-rms" target="_blank">assembly protocol</a> prior to use in CUT&Tag or similar assays (ACT-seq, CoBATCH, TIP-seq and other).</p>
<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
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<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<div class="small-12 medium-12 large-12 columns">B.<br /><center><img alt="pA-Tn5 Transposase unloaded" src="https://www.diagenode.com/img/product/cutandtag/C01070002-fig-b.jpg" /></center></div>
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<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 Transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For flexibility, the fusion protein is not pre-loaded with sequencing adapters. Please follow the recommended <a href="https://www.diagenode.com/en/documents/assembly-protocol-patn5-20042020-rms" target="_blank">assembly protocol</a> prior to use in CUT&Tag or similar assays (ACT-seq, CoBATCH, TIP-seq and other).</p>
<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase" src="https://www.diagenode.com/img/product/cutandtag/C01070002-fig-a.jpg" width="600" height="265" /></center></div>
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<div class="small-12 medium-12 large-12 columns">B.<br /><center><img alt="pA-Tn5 Transposase unloaded" src="https://www.diagenode.com/img/product/cutandtag/C01070002-fig-b.jpg" /></center></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For convenience, the fusion protein is pre-loaded with sequencing adapters. Diagenode pA-Tn5 transposase is compatible with CUT&Tag and other antibody-tethered Tn5 based methods, like ACT-seq and CoBATCH.</p>
<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: loaded or <a href="https://www.diagenode.com/en/p/pa-tn5-transposase-unloaded">unloaded</a><br /></span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag </span></li>
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<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (loaded or <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">unloaded</a>)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
</ul>',
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-a.png" width="800" height="374" /></center><br /> B.<br /><center><img alt="pA-Tn5 Transposase loaded H3K9me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-b.png" width="800" height="374" /></center><br /> C.<br /><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-c.png" width="800" height="374" /></center></div>
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<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1.</strong> Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (A), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">H3K9me3</a> (B) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (C) data obtained using Diagenode pA-Tn5 fusion protein (C01070001) and CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)) at selected loci. CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) and H3K9me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">C15410193</a>).</p>
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<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-a.png" /></center></div>
<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-b.png" /></center></div>
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<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2.</strong> CUT&Tag profiles show typical enrichments specific for a given histone mark. Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (red) and <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (green) are shown at selected loci. H3K27me3, which marks inactive regions (red), does not show enrichment over open chromatin regions, marked by H3K4me3 (green). CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), and H3K27me3 polyclonal ChIP-seq grade (Cat. No.<a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>).</p>
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<div class="small-12 medium-12 large-12 columns">
<div align="center"><video width="400" height="250" autoplay="autoplay" muted="" loop="loop" controls="controls" src="https://www.diagenode.com/videos/chipmentation-dgo.mp4" frameborder="0" allowfullscreen="allowfullscreen"></video></div>
<div align="center"><a href="https://www.diagenode.com/pages/form-chipmentation" class="center alert radius button"><i class="fa fa-info"></i> Contact us</a></div>
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<p>The 24 SI for ChIPmentation includes 24 single primer indexes completing the <a href="https://www.diagenode.com/en/p/tag-kit-for-chipmentation-24">TAG Kit for ChIPmentation</a>. This set of indexes allows for multiplexing up to 24 samples. The TAG Kit for ChIPmentation has been developed for researchers willing to perform ChIPmentation using their own validated ChIP protocol. Our Kit TAG for ChIPmentation includes all reagents necessary for tagmentation-based library preparation compatible with any ChIP protocol based on magnetic beads.</p>
<p>The primer indexes of the 24 SI for ChIPmentation are also compatible with other <strong>tagmentation-based library preparation</strong> <strong>protocols</strong>, such as <strong>ATAC-seq</strong> or <strong>CUT&Tag</strong> technologies.</p>',
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<li>an easier protocol</li>
<li>faster time to results</li>
<li>flexibility – the TAG Kit for ChIPmentation combained with 24 SI for ChIPmentation allows to perform ChIPmentation with <strong>any ChIP protocol </strong>(based on magnetic beads)<strong><br /></strong></li>
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<h3>ChIPmentation on histone marks using TAG KIT for ChIPmentation</h3>
<center><img src="https://www.diagenode.com/img/product/chipmentation/chipmentation-of-histone-using-tag-kit-for-chipmentation.jpg" /></center>
<p><strong>ChIPmentation sequencing results for four histone marks</strong>. Chromatin preparation has been performed on 7 M K562 cells using the Auto iDeal ChIP-seq Kit for Histones (Cat. No. C01010057), TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031)*. Diluted chromatin from 1,000,000 cells was used for the immunoprecipitation with the Diagenode antibodies targeting H3K4me1 (Cat. No. C15410194), H3K27me3 (Cat. No. C15410195), H3K27ac (Cat. No. C15410196) and IgG (Cat. No. C15410206). The experiment was performed on the IP-Star® Compact Automated System.</p>
<p><small>*The kits Auto iDeal ChIP-seq for Histones (C01010057), TAG Kit for ChIPmentation (C01011030) and 24 SI for ChIPmentation (C01011031) form together the Auto ChIPmentation Kit for Histones* (C01011000).</small></p>
<h3>ChIPmentation on non-histone proteins using TAG Kit for ChIPmentation</h3>
<center><img src="https://www.diagenode.com/img/product/chipmentation/chipmentation-of-histone-using-tag-kit-for-chipmentation-2.jpg" /></center>
<p><strong></strong></p>
<p><strong>ChIPmentation sequencing results for two non histone factors</strong>. Chromatin preparation has been performed on 25M K562 cells using the iDeal ChIP-seq kit for TFs (Cat. No. C01010055). Diluted chromatin from 1,000,000 or 4,000,000 cells was used for the immunoprecipitation with the Diagenode antibodies targeting CTCF (Cat. No. C15410210) and RNA PolII (Cat. No. C15100055 ) respectively. The library preparation was performed with the TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031).</p>',
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<p><span><span style="left: 94.4882px; top: 499.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.00223);">The negative Ctrl IgG is intended for use </span><span style="left: 94.4882px; top: 519.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02185);">as a negative control in ChIP, CUT&Tag, MeDIP, IF and other experiments performed with specific antibodies made in rabbit.</span></span></p>',
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<div class="small-8 columns">
<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-if.jpg" alt="Rabbit IgG validated in Immunofluorescence" /></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'description' => '<p><img src="https://www.diagenode.com/img/banners/banner-cutruncuttag-small.png" /></p>
<p><strong>CUT&Tag</strong>-sequencing (<strong>C</strong>leavage <strong>U</strong>nder <strong>T</strong>argets and <strong>Tag</strong>mentation) is a new alternative method to ChIP-seq combining antibody-targeted controlled cleavage by a protein A-Tn5 fusion with massively parallel DNA sequencing to identify the binding sites of DNA-associated proteins. At Diagenode we offer a complete solution for CUT&Tag – our iDeal CUT&Tag for Histones, but also stand-alone fusion protein – pA-Tn5 Transposase. Moreover, we have validated our <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a> in <a href="https://www.diagenode.com/en/categories/cut-and-tag-antibodies">CUT&Tag</a> proving their high performance in this assay.</p>
<ul class="accordion" data-accordion="">
<li class="accordion-navigation"><a href="#v5" style="color: #13b29c;"><i class="fa fa-caret-right"></i> How does it work?</a>
<div id="v5" class="content">
<p>The iDeal CUT&Tag protocol involves the binding of cells on a solid phase ConA magnetic beads, allowing magnetic handling of the cells for the major steps of the protocol. Bead-bound cells are permeabilized, incubated with primary antibody against a target of interest and secondary antibody. Then, Diagenode’s protein pA-Tn5 Transposase - loaded is bound to the complex. Protein A guides Tn5 transposase on chromatin to the antibody attached to its target. Tn5 transposase is activated by Mg+2 ions to insert the sequencing adaptors into genomic regions of interest. DNA is then purified and the tagmented genomic regions of interest are amplified by PCR using Diagenode’s Primer Indexes for tagmented libraries.</p>
<img src="https://www.diagenode.com/img/product/kits/workflow-cutandtag.jpg" /></div>
<h2>Products for CUT&Tag assay</h2>
<h3 class="diacol">Complete solutions</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24" target="_blank">iDeal CUT&Tag kit for Histones</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">Fusion protein</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3064" target="_blank">pA/Tn5 Transposase (loaded)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">pA/Tn5 Transposase (unloaded)</a></li>
</ul>
<h3 class="diacol">CUT&Tag grade antibodies</h3>
<ul class="nobullet">
<li>Antibodies <a href="https://www.diagenode.com/en/applications/cut-and-tag">validated in CUT&Tag</a></li>
<li>Check out our list of <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade antibodies</a></li>
<li>Read more about the performance of Diagenode antibodies in <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">CUT&Tag</a></li>
</ul>
<h3 class="diacol">Positive & Negative CUT&Tag control</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">DNA purification</h3>
<p style="padding-left: 30px;"><a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2<br /></a><a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></p>
<h3 class="diacol">Sequencing indexes</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/primer-indexes-for-tagmented-libraries" target="_blank">Primer indexes for tagmented libraries</a></li>
</ul>
</li>
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<h6 style="height:60px">Rabbit IgG</h6>
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<div class="small-8 columns">
<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</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/C15410206-if.jpg" alt="Rabbit IgG validated in Immunofluorescence" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'description' => '<p>pA-Tn5 transposase is a fusion protein of hyperactive Tn5 transposase and protein A developed for the CUT&Tag assay. For flexibility of use, the fusion protein is not pre-loaded with sequencing adapters. Please follow the recommended assembly protocol prior to use in CUT&Tag or similar assays.</p>',
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'type' => 'Datasheet',
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'date' => '2022-02-01',
'pmid' => 'https://doi.org/10.1101%2F2022.02.17.480825',
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$externalLink = ' <a href="https://doi.org/10.1101%2F2022.02.17.480825" target="_blank"><i class="fa fa-external-link"></i></a>'include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
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Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 200
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
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<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
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<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
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<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
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<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
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<p><strong>pA-Tn5 transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For convenience, the fusion protein is pre-loaded with sequencing adapters. Diagenode pA-Tn5 transposase is compatible with CUT&Tag and other antibody-tethered Tn5 based methods, like ACT-seq and CoBATCH.</p>
<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: loaded or <a href="https://www.diagenode.com/en/p/pa-tn5-transposase-unloaded">unloaded</a><br /></span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag </span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (loaded or <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">unloaded</a>)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
</ul>',
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-a.png" width="800" height="374" /></center><br /> B.<br /><center><img alt="pA-Tn5 Transposase loaded H3K9me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-b.png" width="800" height="374" /></center><br /> C.<br /><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-c.png" width="800" height="374" /></center></div>
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<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1.</strong> Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (A), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">H3K9me3</a> (B) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (C) data obtained using Diagenode pA-Tn5 fusion protein (C01070001) and CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)) at selected loci. CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) and H3K9me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">C15410193</a>).</p>
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<div class="row extra-spaced">
<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-a.png" /></center></div>
<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-b.png" /></center></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2.</strong> CUT&Tag profiles show typical enrichments specific for a given histone mark. Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (red) and <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (green) are shown at selected loci. H3K27me3, which marks inactive regions (red), does not show enrichment over open chromatin regions, marked by H3K4me3 (green). CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), and H3K27me3 polyclonal ChIP-seq grade (Cat. No.<a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>).</p>
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<div class="small-12 medium-12 large-12 columns">
<div align="center"><video width="400" height="250" autoplay="autoplay" muted="" loop="loop" controls="controls" src="https://www.diagenode.com/videos/chipmentation-dgo.mp4" frameborder="0" allowfullscreen="allowfullscreen"></video></div>
<div align="center"><a href="https://www.diagenode.com/pages/form-chipmentation" class="center alert radius button"><i class="fa fa-info"></i> Contact us</a></div>
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<p>The 24 SI for ChIPmentation includes 24 single primer indexes completing the <a href="https://www.diagenode.com/en/p/tag-kit-for-chipmentation-24">TAG Kit for ChIPmentation</a>. This set of indexes allows for multiplexing up to 24 samples. The TAG Kit for ChIPmentation has been developed for researchers willing to perform ChIPmentation using their own validated ChIP protocol. Our Kit TAG for ChIPmentation includes all reagents necessary for tagmentation-based library preparation compatible with any ChIP protocol based on magnetic beads.</p>
<p>The primer indexes of the 24 SI for ChIPmentation are also compatible with other <strong>tagmentation-based library preparation</strong> <strong>protocols</strong>, such as <strong>ATAC-seq</strong> or <strong>CUT&Tag</strong> technologies.</p>',
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<li>an easier protocol</li>
<li>faster time to results</li>
<li>flexibility – the TAG Kit for ChIPmentation combained with 24 SI for ChIPmentation allows to perform ChIPmentation with <strong>any ChIP protocol </strong>(based on magnetic beads)<strong><br /></strong></li>
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<h3>ChIPmentation on histone marks using TAG KIT for ChIPmentation</h3>
<center><img src="https://www.diagenode.com/img/product/chipmentation/chipmentation-of-histone-using-tag-kit-for-chipmentation.jpg" /></center>
<p><strong>ChIPmentation sequencing results for four histone marks</strong>. Chromatin preparation has been performed on 7 M K562 cells using the Auto iDeal ChIP-seq Kit for Histones (Cat. No. C01010057), TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031)*. Diluted chromatin from 1,000,000 cells was used for the immunoprecipitation with the Diagenode antibodies targeting H3K4me1 (Cat. No. C15410194), H3K27me3 (Cat. No. C15410195), H3K27ac (Cat. No. C15410196) and IgG (Cat. No. C15410206). The experiment was performed on the IP-Star® Compact Automated System.</p>
<p><small>*The kits Auto iDeal ChIP-seq for Histones (C01010057), TAG Kit for ChIPmentation (C01011030) and 24 SI for ChIPmentation (C01011031) form together the Auto ChIPmentation Kit for Histones* (C01011000).</small></p>
<h3>ChIPmentation on non-histone proteins using TAG Kit for ChIPmentation</h3>
<center><img src="https://www.diagenode.com/img/product/chipmentation/chipmentation-of-histone-using-tag-kit-for-chipmentation-2.jpg" /></center>
<p><strong></strong></p>
<p><strong>ChIPmentation sequencing results for two non histone factors</strong>. Chromatin preparation has been performed on 25M K562 cells using the iDeal ChIP-seq kit for TFs (Cat. No. C01010055). Diluted chromatin from 1,000,000 or 4,000,000 cells was used for the immunoprecipitation with the Diagenode antibodies targeting CTCF (Cat. No. C15410210) and RNA PolII (Cat. No. C15100055 ) respectively. The library preparation was performed with the TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031).</p>',
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<p><span><span style="left: 94.4882px; top: 499.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.00223);">The negative Ctrl IgG is intended for use </span><span style="left: 94.4882px; top: 519.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02185);">as a negative control in ChIP, CUT&Tag, MeDIP, IF and other experiments performed with specific antibodies made in rabbit.</span></span></p>',
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<div class="small-8 columns">
<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-if.jpg" alt="Rabbit IgG validated in Immunofluorescence" /></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'description' => '<p><img src="https://www.diagenode.com/img/banners/banner-cutruncuttag-small.png" /></p>
<p><strong>CUT&Tag</strong>-sequencing (<strong>C</strong>leavage <strong>U</strong>nder <strong>T</strong>argets and <strong>Tag</strong>mentation) is a new alternative method to ChIP-seq combining antibody-targeted controlled cleavage by a protein A-Tn5 fusion with massively parallel DNA sequencing to identify the binding sites of DNA-associated proteins. At Diagenode we offer a complete solution for CUT&Tag – our iDeal CUT&Tag for Histones, but also stand-alone fusion protein – pA-Tn5 Transposase. Moreover, we have validated our <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a> in <a href="https://www.diagenode.com/en/categories/cut-and-tag-antibodies">CUT&Tag</a> proving their high performance in this assay.</p>
<ul class="accordion" data-accordion="">
<li class="accordion-navigation"><a href="#v5" style="color: #13b29c;"><i class="fa fa-caret-right"></i> How does it work?</a>
<div id="v5" class="content">
<p>The iDeal CUT&Tag protocol involves the binding of cells on a solid phase ConA magnetic beads, allowing magnetic handling of the cells for the major steps of the protocol. Bead-bound cells are permeabilized, incubated with primary antibody against a target of interest and secondary antibody. Then, Diagenode’s protein pA-Tn5 Transposase - loaded is bound to the complex. Protein A guides Tn5 transposase on chromatin to the antibody attached to its target. Tn5 transposase is activated by Mg+2 ions to insert the sequencing adaptors into genomic regions of interest. DNA is then purified and the tagmented genomic regions of interest are amplified by PCR using Diagenode’s Primer Indexes for tagmented libraries.</p>
<img src="https://www.diagenode.com/img/product/kits/workflow-cutandtag.jpg" /></div>
<h2>Products for CUT&Tag assay</h2>
<h3 class="diacol">Complete solutions</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24" target="_blank">iDeal CUT&Tag kit for Histones</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">Fusion protein</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3064" target="_blank">pA/Tn5 Transposase (loaded)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">pA/Tn5 Transposase (unloaded)</a></li>
</ul>
<h3 class="diacol">CUT&Tag grade antibodies</h3>
<ul class="nobullet">
<li>Antibodies <a href="https://www.diagenode.com/en/applications/cut-and-tag">validated in CUT&Tag</a></li>
<li>Check out our list of <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade antibodies</a></li>
<li>Read more about the performance of Diagenode antibodies in <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">CUT&Tag</a></li>
</ul>
<h3 class="diacol">Positive & Negative CUT&Tag control</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">DNA purification</h3>
<p style="padding-left: 30px;"><a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2<br /></a><a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></p>
<h3 class="diacol">Sequencing indexes</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/primer-indexes-for-tagmented-libraries" target="_blank">Primer indexes for tagmented libraries</a></li>
</ul>
</li>
</ul>',
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<h6 style="height:60px">Rabbit IgG</h6>
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<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'name' => 'Spatially resolved epigenomic profiling of single cells in complex tissues',
'authors' => 'Lu Tian et al.',
'description' => '<p>The recent development of spatial omics methods enables single-cell profiling of the transcriptome and the 3D genome organization in a spatially resolved manner. Expanding the repertoire of spatial omics tools, a spatial epigenomics method will accelerate our understanding of the spatial regulation of cell and tissue functions. Here, we report a method for spatially resolved profiling of epigenomes in single cells using in-situ tagmentation and transcription followed by highly multiplexed imaging. We profiled histone modifications marking active promoters and enhancers, H3K4me3 and H3K27ac, and generated high-resolution spatial atlas of hundreds of active promoters and putative enhancers in embryonic and adult mouse brains. Our results further revealed putative promoter-enhancer pairs and enhancer hubs regulating the expression of developmentally important genes. We envision this approach will be generally applicable to spatial profiling of epigenetic modifications and DNA-binding proteins, advancing our understanding of how gene expression is spatiotemporally regulated by the epigenome.</p>',
'date' => '2022-02-01',
'pmid' => 'https://doi.org/10.1101%2F2022.02.17.480825',
'doi' => '10.1101/2022.02.17.480825',
'modified' => '2022-08-11 15:16:56',
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<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
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<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
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<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
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<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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'meta_description' => 'pA-Tn5 Transposase (unloaded) is a fusion protein of hyperactive Tn5 transposase and protein A. For flexibility, the fusion protein is not pre-loaded with sequencing adapters.',
'modified' => '2022-11-25 15:00:26',
'created' => '2020-04-28 16:21:20'
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'description' => '<p><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24"> <img alt="pA-Tn5 Transposase loaded" src="https://www.diagenode.com/img/banners/banner-cutruncuttag-small.png" /></a></p>
<p><strong>pA-Tn5 transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For convenience, the fusion protein is pre-loaded with sequencing adapters. Diagenode pA-Tn5 transposase is compatible with CUT&Tag and other antibody-tethered Tn5 based methods, like ACT-seq and CoBATCH.</p>
<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: loaded or <a href="https://www.diagenode.com/en/p/pa-tn5-transposase-unloaded">unloaded</a><br /></span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag </span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (loaded or <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">unloaded</a>)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
</ul>',
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-a.png" width="800" height="374" /></center><br /> B.<br /><center><img alt="pA-Tn5 Transposase loaded H3K9me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-b.png" width="800" height="374" /></center><br /> C.<br /><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-c.png" width="800" height="374" /></center></div>
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<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1.</strong> Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (A), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">H3K9me3</a> (B) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (C) data obtained using Diagenode pA-Tn5 fusion protein (C01070001) and CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)) at selected loci. CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) and H3K9me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">C15410193</a>).</p>
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<div class="row extra-spaced">
<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-a.png" /></center></div>
<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-b.png" /></center></div>
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<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2.</strong> CUT&Tag profiles show typical enrichments specific for a given histone mark. Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (red) and <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (green) are shown at selected loci. H3K27me3, which marks inactive regions (red), does not show enrichment over open chromatin regions, marked by H3K4me3 (green). CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), and H3K27me3 polyclonal ChIP-seq grade (Cat. No.<a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>).</p>
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'meta_description' => 'Diagenode pA-Tn5 Transposase (loaded) is a fusion protein of hyperactive Tn5 transposase and protein A developed for the CUT&Tag assay. For convenience, the fusion protein is pre-loaded with sequencing adapters.',
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<div class="small-12 medium-12 large-12 columns">
<div align="center"><video width="400" height="250" autoplay="autoplay" muted="" loop="loop" controls="controls" src="https://www.diagenode.com/videos/chipmentation-dgo.mp4" frameborder="0" allowfullscreen="allowfullscreen"></video></div>
<div align="center"><a href="https://www.diagenode.com/pages/form-chipmentation" class="center alert radius button"><i class="fa fa-info"></i> Contact us</a></div>
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<p>The 24 SI for ChIPmentation includes 24 single primer indexes completing the <a href="https://www.diagenode.com/en/p/tag-kit-for-chipmentation-24">TAG Kit for ChIPmentation</a>. This set of indexes allows for multiplexing up to 24 samples. The TAG Kit for ChIPmentation has been developed for researchers willing to perform ChIPmentation using their own validated ChIP protocol. Our Kit TAG for ChIPmentation includes all reagents necessary for tagmentation-based library preparation compatible with any ChIP protocol based on magnetic beads.</p>
<p>The primer indexes of the 24 SI for ChIPmentation are also compatible with other <strong>tagmentation-based library preparation</strong> <strong>protocols</strong>, such as <strong>ATAC-seq</strong> or <strong>CUT&Tag</strong> technologies.</p>',
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'info1' => '<h3>Comparing to classical ChIP-seq, ChIPmentation enables:</h3>
<ul>
<li>an easier protocol</li>
<li>faster time to results</li>
<li>flexibility – the TAG Kit for ChIPmentation combained with 24 SI for ChIPmentation allows to perform ChIPmentation with <strong>any ChIP protocol </strong>(based on magnetic beads)<strong><br /></strong></li>
</ul>
<h3>ChIPmentation on histone marks using TAG KIT for ChIPmentation</h3>
<center><img src="https://www.diagenode.com/img/product/chipmentation/chipmentation-of-histone-using-tag-kit-for-chipmentation.jpg" /></center>
<p><strong>ChIPmentation sequencing results for four histone marks</strong>. Chromatin preparation has been performed on 7 M K562 cells using the Auto iDeal ChIP-seq Kit for Histones (Cat. No. C01010057), TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031)*. Diluted chromatin from 1,000,000 cells was used for the immunoprecipitation with the Diagenode antibodies targeting H3K4me1 (Cat. No. C15410194), H3K27me3 (Cat. No. C15410195), H3K27ac (Cat. No. C15410196) and IgG (Cat. No. C15410206). The experiment was performed on the IP-Star® Compact Automated System.</p>
<p><small>*The kits Auto iDeal ChIP-seq for Histones (C01010057), TAG Kit for ChIPmentation (C01011030) and 24 SI for ChIPmentation (C01011031) form together the Auto ChIPmentation Kit for Histones* (C01011000).</small></p>
<h3>ChIPmentation on non-histone proteins using TAG Kit for ChIPmentation</h3>
<center><img src="https://www.diagenode.com/img/product/chipmentation/chipmentation-of-histone-using-tag-kit-for-chipmentation-2.jpg" /></center>
<p><strong></strong></p>
<p><strong>ChIPmentation sequencing results for two non histone factors</strong>. Chromatin preparation has been performed on 25M K562 cells using the iDeal ChIP-seq kit for TFs (Cat. No. C01010055). Diluted chromatin from 1,000,000 or 4,000,000 cells was used for the immunoprecipitation with the Diagenode antibodies targeting CTCF (Cat. No. C15410210) and RNA PolII (Cat. No. C15100055 ) respectively. The library preparation was performed with the TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031).</p>',
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'description' => '<p><span>The negative Ctrl IgG from rabbit has been extensively validated in chromatin immunoprecipitation assays (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy animals. This IgG preparation is intended for use as a negative control in ChIP experiments for specific antibodies made in rabbit. The negative Ctrl IgG from rabbit should be used for ChIP in parallel with specific antibody at the same concentration as the specific antibody. </span></p>
<p><span><span style="left: 94.4882px; top: 499.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.00223);">The negative Ctrl IgG is intended for use </span><span style="left: 94.4882px; top: 519.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02185);">as a negative control in ChIP, CUT&Tag, MeDIP, IF and other experiments performed with specific antibodies made in rabbit.</span></span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-chip.jpg" alt="Rabbit IgG Antibody ChIP Grade" /></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-if.jpg" alt="Rabbit IgG validated in Immunofluorescence" /></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'info2' => '<p>The negative control IgG from rabbit has been extensively validated in chromatin immunoprecipitation (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy rabbits. This IgG preparation is intended for use as a negative control in ChIP, MeDIP, IF and other experiments performed with specific antibodies made in rabbit. The negative control IgG from rabbit should be used in parallel with the specific antibody at the same concentration. It is also included in many of our ChIP and MeDIP kits.</p>',
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'id' => '142',
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'description' => '<p><img src="https://www.diagenode.com/img/banners/banner-cutruncuttag-small.png" /></p>
<p><strong>CUT&Tag</strong>-sequencing (<strong>C</strong>leavage <strong>U</strong>nder <strong>T</strong>argets and <strong>Tag</strong>mentation) is a new alternative method to ChIP-seq combining antibody-targeted controlled cleavage by a protein A-Tn5 fusion with massively parallel DNA sequencing to identify the binding sites of DNA-associated proteins. At Diagenode we offer a complete solution for CUT&Tag – our iDeal CUT&Tag for Histones, but also stand-alone fusion protein – pA-Tn5 Transposase. Moreover, we have validated our <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a> in <a href="https://www.diagenode.com/en/categories/cut-and-tag-antibodies">CUT&Tag</a> proving their high performance in this assay.</p>
<ul class="accordion" data-accordion="">
<li class="accordion-navigation"><a href="#v5" style="color: #13b29c;"><i class="fa fa-caret-right"></i> How does it work?</a>
<div id="v5" class="content">
<p>The iDeal CUT&Tag protocol involves the binding of cells on a solid phase ConA magnetic beads, allowing magnetic handling of the cells for the major steps of the protocol. Bead-bound cells are permeabilized, incubated with primary antibody against a target of interest and secondary antibody. Then, Diagenode’s protein pA-Tn5 Transposase - loaded is bound to the complex. Protein A guides Tn5 transposase on chromatin to the antibody attached to its target. Tn5 transposase is activated by Mg+2 ions to insert the sequencing adaptors into genomic regions of interest. DNA is then purified and the tagmented genomic regions of interest are amplified by PCR using Diagenode’s Primer Indexes for tagmented libraries.</p>
<img src="https://www.diagenode.com/img/product/kits/workflow-cutandtag.jpg" /></div>
<h2>Products for CUT&Tag assay</h2>
<h3 class="diacol">Complete solutions</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24" target="_blank">iDeal CUT&Tag kit for Histones</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">Fusion protein</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3064" target="_blank">pA/Tn5 Transposase (loaded)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">pA/Tn5 Transposase (unloaded)</a></li>
</ul>
<h3 class="diacol">CUT&Tag grade antibodies</h3>
<ul class="nobullet">
<li>Antibodies <a href="https://www.diagenode.com/en/applications/cut-and-tag">validated in CUT&Tag</a></li>
<li>Check out our list of <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade antibodies</a></li>
<li>Read more about the performance of Diagenode antibodies in <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">CUT&Tag</a></li>
</ul>
<h3 class="diacol">Positive & Negative CUT&Tag control</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">DNA purification</h3>
<p style="padding-left: 30px;"><a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2<br /></a><a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></p>
<h3 class="diacol">Sequencing indexes</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/primer-indexes-for-tagmented-libraries" target="_blank">Primer indexes for tagmented libraries</a></li>
</ul>
</li>
</ul>',
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<h6 style="height:60px">Rabbit IgG</h6>
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<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'description' => '<p>pA-Tn5 transposase is a fusion protein of hyperactive Tn5 transposase and protein A developed for the CUT&Tag assay. For flexibility of use, the fusion protein is not pre-loaded with sequencing adapters. Please follow the recommended assembly protocol prior to use in CUT&Tag or similar assays.</p>',
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'name' => 'Spatially resolved epigenomic profiling of single cells in complex tissues',
'authors' => 'Lu Tian et al.',
'description' => '<p>The recent development of spatial omics methods enables single-cell profiling of the transcriptome and the 3D genome organization in a spatially resolved manner. Expanding the repertoire of spatial omics tools, a spatial epigenomics method will accelerate our understanding of the spatial regulation of cell and tissue functions. Here, we report a method for spatially resolved profiling of epigenomes in single cells using in-situ tagmentation and transcription followed by highly multiplexed imaging. We profiled histone modifications marking active promoters and enhancers, H3K4me3 and H3K27ac, and generated high-resolution spatial atlas of hundreds of active promoters and putative enhancers in embryonic and adult mouse brains. Our results further revealed putative promoter-enhancer pairs and enhancer hubs regulating the expression of developmentally important genes. We envision this approach will be generally applicable to spatial profiling of epigenetic modifications and DNA-binding proteins, advancing our understanding of how gene expression is spatiotemporally regulated by the epigenome.</p>',
'date' => '2022-02-01',
'pmid' => 'https://doi.org/10.1101%2F2022.02.17.480825',
'doi' => '10.1101/2022.02.17.480825',
'modified' => '2022-08-11 15:16:56',
'created' => '2022-08-11 12:14:50',
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<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
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<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
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<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
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<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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'slug' => 'pa-tn5-transposase-unloaded',
'meta_title' => 'pA-Tn5 Transposase (unloaded) | Diagenode',
'meta_keywords' => '',
'meta_description' => 'pA-Tn5 Transposase (unloaded) is a fusion protein of hyperactive Tn5 transposase and protein A. For flexibility, the fusion protein is not pre-loaded with sequencing adapters.',
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'description' => '<p><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24"> <img alt="pA-Tn5 Transposase loaded" src="https://www.diagenode.com/img/banners/banner-cutruncuttag-small.png" /></a></p>
<p><strong>pA-Tn5 transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For convenience, the fusion protein is pre-loaded with sequencing adapters. Diagenode pA-Tn5 transposase is compatible with CUT&Tag and other antibody-tethered Tn5 based methods, like ACT-seq and CoBATCH.</p>
<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: loaded or <a href="https://www.diagenode.com/en/p/pa-tn5-transposase-unloaded">unloaded</a><br /></span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag </span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a></li>
<li>pA/Tn5 Transposase (loaded or <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">unloaded</a>)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
</ul>',
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-a.png" width="800" height="374" /></center><br /> B.<br /><center><img alt="pA-Tn5 Transposase loaded H3K9me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-b.png" width="800" height="374" /></center><br /> C.<br /><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-c.png" width="800" height="374" /></center></div>
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<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1.</strong> Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (A), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">H3K9me3</a> (B) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (C) data obtained using Diagenode pA-Tn5 fusion protein (C01070001) and CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)) at selected loci. CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) and H3K9me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">C15410193</a>).</p>
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<div class="row extra-spaced">
<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-a.png" /></center></div>
<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-b.png" /></center></div>
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<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2.</strong> CUT&Tag profiles show typical enrichments specific for a given histone mark. Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (red) and <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (green) are shown at selected loci. H3K27me3, which marks inactive regions (red), does not show enrichment over open chromatin regions, marked by H3K4me3 (green). CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), and H3K27me3 polyclonal ChIP-seq grade (Cat. No.<a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>).</p>
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'meta_description' => 'Diagenode pA-Tn5 Transposase (loaded) is a fusion protein of hyperactive Tn5 transposase and protein A developed for the CUT&Tag assay. For convenience, the fusion protein is pre-loaded with sequencing adapters.',
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<div class="small-12 medium-12 large-12 columns">
<div align="center"><video width="400" height="250" autoplay="autoplay" muted="" loop="loop" controls="controls" src="https://www.diagenode.com/videos/chipmentation-dgo.mp4" frameborder="0" allowfullscreen="allowfullscreen"></video></div>
<div align="center"><a href="https://www.diagenode.com/pages/form-chipmentation" class="center alert radius button"><i class="fa fa-info"></i> Contact us</a></div>
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<p>The 24 SI for ChIPmentation includes 24 single primer indexes completing the <a href="https://www.diagenode.com/en/p/tag-kit-for-chipmentation-24">TAG Kit for ChIPmentation</a>. This set of indexes allows for multiplexing up to 24 samples. The TAG Kit for ChIPmentation has been developed for researchers willing to perform ChIPmentation using their own validated ChIP protocol. Our Kit TAG for ChIPmentation includes all reagents necessary for tagmentation-based library preparation compatible with any ChIP protocol based on magnetic beads.</p>
<p>The primer indexes of the 24 SI for ChIPmentation are also compatible with other <strong>tagmentation-based library preparation</strong> <strong>protocols</strong>, such as <strong>ATAC-seq</strong> or <strong>CUT&Tag</strong> technologies.</p>',
'label1' => 'Characteristics',
'info1' => '<h3>Comparing to classical ChIP-seq, ChIPmentation enables:</h3>
<ul>
<li>an easier protocol</li>
<li>faster time to results</li>
<li>flexibility – the TAG Kit for ChIPmentation combained with 24 SI for ChIPmentation allows to perform ChIPmentation with <strong>any ChIP protocol </strong>(based on magnetic beads)<strong><br /></strong></li>
</ul>
<h3>ChIPmentation on histone marks using TAG KIT for ChIPmentation</h3>
<center><img src="https://www.diagenode.com/img/product/chipmentation/chipmentation-of-histone-using-tag-kit-for-chipmentation.jpg" /></center>
<p><strong>ChIPmentation sequencing results for four histone marks</strong>. Chromatin preparation has been performed on 7 M K562 cells using the Auto iDeal ChIP-seq Kit for Histones (Cat. No. C01010057), TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031)*. Diluted chromatin from 1,000,000 cells was used for the immunoprecipitation with the Diagenode antibodies targeting H3K4me1 (Cat. No. C15410194), H3K27me3 (Cat. No. C15410195), H3K27ac (Cat. No. C15410196) and IgG (Cat. No. C15410206). The experiment was performed on the IP-Star® Compact Automated System.</p>
<p><small>*The kits Auto iDeal ChIP-seq for Histones (C01010057), TAG Kit for ChIPmentation (C01011030) and 24 SI for ChIPmentation (C01011031) form together the Auto ChIPmentation Kit for Histones* (C01011000).</small></p>
<h3>ChIPmentation on non-histone proteins using TAG Kit for ChIPmentation</h3>
<center><img src="https://www.diagenode.com/img/product/chipmentation/chipmentation-of-histone-using-tag-kit-for-chipmentation-2.jpg" /></center>
<p><strong></strong></p>
<p><strong>ChIPmentation sequencing results for two non histone factors</strong>. Chromatin preparation has been performed on 25M K562 cells using the iDeal ChIP-seq kit for TFs (Cat. No. C01010055). Diluted chromatin from 1,000,000 or 4,000,000 cells was used for the immunoprecipitation with the Diagenode antibodies targeting CTCF (Cat. No. C15410210) and RNA PolII (Cat. No. C15100055 ) respectively. The library preparation was performed with the TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031).</p>',
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'meta_description' => 'Diagenode 24 Single indexes for ChIPmentation are compatible with any tagmentation-based library prepration protocols, such as ChIPmentation, ATAC-seq or CUT&Tag technologies. This set of indexes allows for multiplexing up to 24 samples.',
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'description' => '<p><span>The negative Ctrl IgG from rabbit has been extensively validated in chromatin immunoprecipitation assays (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy animals. This IgG preparation is intended for use as a negative control in ChIP experiments for specific antibodies made in rabbit. The negative Ctrl IgG from rabbit should be used for ChIP in parallel with specific antibody at the same concentration as the specific antibody. </span></p>
<p><span><span style="left: 94.4882px; top: 499.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.00223);">The negative Ctrl IgG is intended for use </span><span style="left: 94.4882px; top: 519.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02185);">as a negative control in ChIP, CUT&Tag, MeDIP, IF and other experiments performed with specific antibodies made in rabbit.</span></span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-chip.jpg" alt="Rabbit IgG Antibody ChIP Grade" /></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-if.jpg" alt="Rabbit IgG validated in Immunofluorescence" /></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'info2' => '<p>The negative control IgG from rabbit has been extensively validated in chromatin immunoprecipitation (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy rabbits. This IgG preparation is intended for use as a negative control in ChIP, MeDIP, IF and other experiments performed with specific antibodies made in rabbit. The negative control IgG from rabbit should be used in parallel with the specific antibody at the same concentration. It is also included in many of our ChIP and MeDIP kits.</p>',
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'id' => '142',
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'name' => 'CUT&Tag',
'description' => '<p><img src="https://www.diagenode.com/img/banners/banner-cutruncuttag-small.png" /></p>
<p><strong>CUT&Tag</strong>-sequencing (<strong>C</strong>leavage <strong>U</strong>nder <strong>T</strong>argets and <strong>Tag</strong>mentation) is a new alternative method to ChIP-seq combining antibody-targeted controlled cleavage by a protein A-Tn5 fusion with massively parallel DNA sequencing to identify the binding sites of DNA-associated proteins. At Diagenode we offer a complete solution for CUT&Tag – our iDeal CUT&Tag for Histones, but also stand-alone fusion protein – pA-Tn5 Transposase. Moreover, we have validated our <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a> in <a href="https://www.diagenode.com/en/categories/cut-and-tag-antibodies">CUT&Tag</a> proving their high performance in this assay.</p>
<ul class="accordion" data-accordion="">
<li class="accordion-navigation"><a href="#v5" style="color: #13b29c;"><i class="fa fa-caret-right"></i> How does it work?</a>
<div id="v5" class="content">
<p>The iDeal CUT&Tag protocol involves the binding of cells on a solid phase ConA magnetic beads, allowing magnetic handling of the cells for the major steps of the protocol. Bead-bound cells are permeabilized, incubated with primary antibody against a target of interest and secondary antibody. Then, Diagenode’s protein pA-Tn5 Transposase - loaded is bound to the complex. Protein A guides Tn5 transposase on chromatin to the antibody attached to its target. Tn5 transposase is activated by Mg+2 ions to insert the sequencing adaptors into genomic regions of interest. DNA is then purified and the tagmented genomic regions of interest are amplified by PCR using Diagenode’s Primer Indexes for tagmented libraries.</p>
<img src="https://www.diagenode.com/img/product/kits/workflow-cutandtag.jpg" /></div>
<h2>Products for CUT&Tag assay</h2>
<h3 class="diacol">Complete solutions</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24" target="_blank">iDeal CUT&Tag kit for Histones</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">Fusion protein</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3064" target="_blank">pA/Tn5 Transposase (loaded)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">pA/Tn5 Transposase (unloaded)</a></li>
</ul>
<h3 class="diacol">CUT&Tag grade antibodies</h3>
<ul class="nobullet">
<li>Antibodies <a href="https://www.diagenode.com/en/applications/cut-and-tag">validated in CUT&Tag</a></li>
<li>Check out our list of <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade antibodies</a></li>
<li>Read more about the performance of Diagenode antibodies in <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">CUT&Tag</a></li>
</ul>
<h3 class="diacol">Positive & Negative CUT&Tag control</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">DNA purification</h3>
<p style="padding-left: 30px;"><a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2<br /></a><a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></p>
<h3 class="diacol">Sequencing indexes</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/primer-indexes-for-tagmented-libraries" target="_blank">Primer indexes for tagmented libraries</a></li>
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<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'id' => '1079',
'name' => 'pA-TN5 transposase (unloaded) datasheet',
'description' => '<p>pA-Tn5 transposase is a fusion protein of hyperactive Tn5 transposase and protein A developed for the CUT&Tag assay. For flexibility of use, the fusion protein is not pre-loaded with sequencing adapters. Please follow the recommended assembly protocol prior to use in CUT&Tag or similar assays.</p>',
'image_id' => null,
'type' => 'Datasheet',
'url' => 'files/products/cutandtag/C01070002-pa-tn5-transposase-unloaded.pdf',
'slug' => 'c01070002-pa-tn5-transposase-unloaded-tds',
'meta_keywords' => '',
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'modified' => '2020-04-28 16:45:02',
'created' => '2020-04-28 16:45:02',
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'id' => '4408',
'name' => 'Spatially resolved epigenomic profiling of single cells in complex tissues',
'authors' => 'Lu Tian et al.',
'description' => '<p>The recent development of spatial omics methods enables single-cell profiling of the transcriptome and the 3D genome organization in a spatially resolved manner. Expanding the repertoire of spatial omics tools, a spatial epigenomics method will accelerate our understanding of the spatial regulation of cell and tissue functions. Here, we report a method for spatially resolved profiling of epigenomes in single cells using in-situ tagmentation and transcription followed by highly multiplexed imaging. We profiled histone modifications marking active promoters and enhancers, H3K4me3 and H3K27ac, and generated high-resolution spatial atlas of hundreds of active promoters and putative enhancers in embryonic and adult mouse brains. Our results further revealed putative promoter-enhancer pairs and enhancer hubs regulating the expression of developmentally important genes. We envision this approach will be generally applicable to spatial profiling of epigenetic modifications and DNA-binding proteins, advancing our understanding of how gene expression is spatiotemporally regulated by the epigenome.</p>',
'date' => '2022-02-01',
'pmid' => 'https://doi.org/10.1101%2F2022.02.17.480825',
'doi' => '10.1101/2022.02.17.480825',
'modified' => '2022-08-11 15:16:56',
'created' => '2022-08-11 12:14:50',
'ProductsPublication' => array(
'id' => '6030',
'product_id' => '3065',
'publication_id' => '4408'
)
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