GR Antibody - ChIP-seq Grade (C15200010)

Lot	NR-010
Concentration	1.0 µg/µl
Species reactivity	Human, rat
Type	Monoclonal
Purity	Ammonium sulphate purified
Host	Mouse
Precautions	This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Applications	Suggested dilution	References
ChIP/ChIP-seq	5 μg/ChIP	Fig 1, 2
ELISA	0.5 μg/ml	Fig 3
Western Blotting	1 μg/ml	Fig 4
Gel Supershift	5 μg/ml
Immunochemistry	2.5 μg/ml	Fig 5, 6
Flow cytometry	0.5 μg/ml
Immunoprecipitation	5 μg/IP	Fig 7

GR Antibody ChIP Grade

Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against hGR
ChIP assays were performed using HeLa cells, the Diagenode monoclonal antibody directed against GR (Cat. No. MAb-010-050) and optimized PCR primer sets for qPCR. The cells were treated either with ethanol (EtOH, used as a negative control) or triamcinolone acetonide (TA) for 4 hours prior to cell harvesting. ChIP was performed using sheared chromatin from 3 million cells and 5 μg of antibody. QPCR was performed with primers for the human metallothionein promoter (hMTIIA) and for exon 2 of the human myoglobin gene (hmyo ex2), used as a negative control. Figure 1 shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +/- control target). These results demonstrate the occupancy of the human metallothionein IIA promoter by GR.

GR Antibody ChIP-seq Grade

GR Antibody for ChIP-seq

GR Antibody for ChIP-seq assay

GR Antibody validated in ChIP-seq

GR Antibody ChIP-seq Grade

Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against hGR
ChIP was performed on sheared chromatin from 3.5 million HeLaB2 cells using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). The cells were treated with the synthetic GR ligand triamcinolone acetonide (TA) for 4 hours prior to harvesting. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence of chromosome 16 (figure 2A) as well as the MT2A positive control gene (figure 2B). The position of the PCR amplicon is also indicated. Figure 2C, D and E show the results for the known GR target genes PER1 on chromosome 17 and FKBP5 and TNFAIP3 on chromosome 6.

GR Antibody ELISA Validation

Figure 3. Sandwich ELISA
The specificity of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050) was assessed by sandwich ELISA. Figure 3A: schematic representation of the sandwich ELISA with the monoclonal antibody against hGR (clone #:m2F8). Figure 3B: ELISA results using the monoclonal antibody against hGR at a concentration of 0.5 μg/ml. The figure shows an ELISA signal which is proportionally increasing with increasing amounts of recombinant hGR.

GR Antibody validated in Western Blot

Figure 4. Western blot analysis using the Diagenode monoclonal antibody against hGR
Figure 4A. Extracts from HeLa cells containing the indicated amounts of GR (from 30 to 240 fmol), and from 5x10e6 Raji or Molt cells were analysed by Western blot using the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050). Figure 4B. Western blot analysis of extracts from 300,000 HeLa cells with the Diagenode monoclonal antibody against hGR (concentration 1 μg/ml).

GR Antibody validated in Immunohistochemistry

GR Antibody validated for Immunohistochemistry

Figure 5. Immunohistochemistry and immunofluorescence using the Diagenode monoclonal antibody against hGR
1. Immunoreactivity of the Diagenode monoclonal antibody against hGR (Cat. No. MAb-010-050) in rat CA1 neurons of hippocampus. The antibody was used at a concentration of 2.5 μg/ml.
2. COS-7 cells transiently overexpressing human GR were labeled with the antibody against hGR followed by a biotinylated secondary antibody and peroxidase-labeled avidin. The antibody was used at a concentration of 2.5 μg/ml.

GR Antibody validated in Immunoprecipitation

Figure 6. Immunoprecipitation using the Diagenode monoclonal antibody against hGR
The glucocorticoid receptor was immunoprecipitated from HeLa cell extracts (5 million HeLa cells in 100 μl IP reaction solution) using 5 μg of the Diagenode monoclonal antibody directed against hGR (Cat. No. MAb-010-050). The IP was followed by Western blot analysis as described above

Target Description

Monoclonal antibody raised in mouse against amino acids 304-428 of the human GR (glucocorticoid receptor), using a chimeric protein.

Applications

ELISA
Enzyme-linked immunosorbent assay. Read more

WB
Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more

GSA
Gel Supershift Read more

Flow Cyt
Flow cyt Read more

IP
Immunoprecipitation Read more

ChIP-seq (ab)
Read more

ChIP-qPCR (ab)
Read more

Documents

Datasheet hGR C15200010 DATASHEET Datasheet description	Download
Antibodies you can trust POSTER Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...	Download
Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...	Download

Safety sheets

GR monoclonal antibody GB en	Download
GR monoclonal antibody US en	Download
GR monoclonal antibody JP ja	Download
GR monoclonal antibody SDS DE de	Download
GR monoclonal antibody SDS BE nl	Download
GR monoclonal antibody SDS FR fr	Download
GR monoclonal antibody SDS BE fr	Download
GR monoclonal antibody SDS ES es	Download

Publications

How to properly cite this product in your work

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The Role of S-Palmitoylation of the Human Glucocorticoid Receptor (hGR) in Mediating the Nongenomic Glucocorticoid Actions
Nicolaides N. et al.
Background: Many rapid nongenomic glucocorticoid actions are mediated by membrane-bound glucocorticoid receptors (GRs). S-palmitoylation is a lipid post-translational modification that mediates the membrane localization of some steroid receptors. A highly homologous amino acid sequence (663YLCM KTLLL671) is present ...

Three Novel Heterozygous Point Mutations of NR3C1 causing Glucocorticoid Resistance
Vitellius G et al.
Generalized glucocorticoid resistance is associated with glucocorticoid receptor (GR, NR3C1) mutations. Three novel heterozygous missense NR3C1 mutations (R477S, Y478C and L672P) were identified in patients presenting with adrenal incidentalomas, glucocorticoid excess without Cushing syndrome. Dexamethasone (DXM) bi...

Functional characterization of the hGRαT556I causing Chrousos syndrome
Nicolaides NC et al.
BACKGROUND: Chrousos syndrome is a rare pathologic condition characterized by generalized, partial resistance of target tissues to glucocorticoids and caused by inactivating mutations of the human glucocorticoid receptor (hGR) gene. A novel case of Chrousos syndrome has been reported in a patient with adrenal incid...

Transient generalized glucocorticoid hypersensitivity
Nicolaides NC et al.
BACKGROUND: Transient generalized glucocorticoid hypersensitivity is a rare disorder characterized by increased tissue sensitivity to glucocorticoids and compensatory hypo-activation of the hypothalamic-pituitary-adrenal axis. The condition itself and the underlying molecular mechanisms have not been elucidated. O...

Perinatal exposure to low-dose bisphenol A affects the neuroendocrine stress response in rats.
Panagiotidou E, Zerva S, Mitsiou DJ, Alexis MN, Kitraki E
Bisphenol A (BPA) is an estrogen-mimicking endocrine disruptor. Early-life exposures to low doses of BPA exert long-lasting effects on animals' reproductive and brain physiology. However, little is known about the effects of BPA on the stress-response system. Given the interaction of sex and stress hormones, we exam...

Pattern of heat shock factor and heat shock protein expression in lymphocytes of bipolar patients: Increased HSP70-glucocorticoid receptor heterocomplex.
Bei ES, Salpeas V, Alevizos B, Anagnostara C, Pappa D, Moutsatsou P
Bipolar disorder (BD), a stress-related disease, is characterized by altered glucocorticoid receptor (GR) signalling. Stress response includes activation of heat shock factor (HSF) and subsequent heat shock protein (HSP) synthesis which regulate GR folding and function. The objective of this study was to investigate...

Coactivation of GR and NFKB alters the repertoire of their binding sites and target genes.
Rao NA, McCalman MT, Moulos P, Francoijs KJ, Chatziioannou A, Kolisis FN, Alexis MN, Mitsiou DJ, Stunnenberg HG
Glucocorticoid receptor (GR) exerts anti-inflammatory action in part by antagonizing proinflammatory transcription factors such as the nuclear factor kappa-b (NFKB). Here, we assess the crosstalk of activated GR and RELA (p65, major NFKB component) by global identification of their binding sites and target genes. We...

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GR Antibody - ChIP-seq Grade

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