Anti-Glucocorticoid Receptor antibody [BuGR2] - ChIP Grade (ab2768)

Overview

  • Product nameAnti-Glucocorticoid Receptor antibody [BuGR2] - ChIP Grade
    See all Glucocorticoid Receptor primary antibodies
  • Description
    Mouse monoclonal [BuGR2] to Glucocorticoid Receptor - ChIP Grade
  • SpecificityImmunocytochemical staining of GR in L929 cells with this antibody results in staining of both the cytoplasm and nucleus, even in the presence of hormone. This antibody, using enzymatic digestion analysis, has been shown to react with the undigested 97 kDa GR, a 17 kDa DNA-binding trypsin fragment, and a 45 kDa steroid- and DNA-binding chymotrypsin fragment.
  • Tested applicationsSuitable for: ICC/IF, ChIP, ELISA, IHC-FoFr, Flow Cyt, IHC-P, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Rabbit, Guinea pig, Human, Saccharomyces cerevisiae, Xenopus laevis
    Does not react with: Bird, Non Human Primates, Amphibians
  • Immunogen

    Other Immunogen Type corresponding to Rat Glucocorticoid Receptor. Partially purified rat GR.

  • Positive control
    • ICC: L929 cells WB: Mouse pituitary tumors, mammary tissue, placenta, liver and thymus lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab2768 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/50 - 1/500.
ChIP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC-FoFr 1/500. PubMed: 20307510
EMSA Use at an assay dependent concentration.
Flow Cyt Use 0.5-1µg for 106 cells.

ab18414 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IHC-P Use a concentration of 5 µg/ml. We recommend the following for antigen retrieval: Incubate in 10 mM citrate buffer, pH 6.0 in microwave oven for 7 x 2 minutes (2 minutes, stop, 2 minutes, etc.). Then wash with Dulbecco’s PBS 6 x 7 minutes at room temperature. The tissue is now ready to be blocked for the start of IHC.
IP Use at an assay dependent concentration.
WB Use a concentration of 5 µg/ml. Detects a band of approximately 97 kDa (predicted molecular weight: 86 kDa). Using enzymatic digestion analysis detects a band of approximately 97 kDa, a 17 kDa DNA-binding trypsin fragment, and a 45 kDa steroid- and DNA-binding chymotrypsin fragment (predicted molecular weight: 86 kDa).

Target

  • FunctionReceptor for glucocorticoids (GC). Has a dual mode of action: as a transcription factor that binds to glucocorticoid response elements (GRE) and as a modulator of other transcription factors. Affects inflammatory responses, cellular proliferation and differentiation in target tissues. Could act as a coactivator for STAT5-dependent transcription upon growth hormone (GH) stimulation and could reveal an essential role of hepatic GR in the control of body growth. Involved in chromatin remodeling. Plays a significant role in transactivation. Involved in nuclear translocation.
  • Tissue specificityWidely expressed. In the heart, detected in left and right atria, left and right ventricles, aorta, apex, intraventricular septum, and atrioventricular node as well as whole adult and fetal heart.
  • Involvement in diseaseDefects in NR3C1 are a cause of glucocorticoid resistance (GCRES) [MIM:138040]; also known as cortisol resistance. It is a hypertensive, hyperandrogenic disorder characterized by increased serum cortisol concentrations. Inheritance is autosomal dominant.
  • Sequence similaritiesBelongs to the nuclear hormone receptor family. NR3 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • DomainComposed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
  • Post-translational
    modifications
    Increased proteasome-mediated degradation in response to glucocorticoids.
    Phosphorylated in the absence of hormone; becomes hyperphosphorylated in the presence of glucocorticoid. The Ser-203-phosphorylated form is mainly cytoplasmic, and the Ser-211-phosphorylated form is nuclear. Transcriptional activity correlates with the amount of phosphorylation at Ser-211.
    Sumoylated; this reduces transcription transactivation.
    Ubiquitinated; restricts glucocorticoid-mediated transcriptional signaling.
  • Cellular localizationCytoplasm. Nucleus. Cytoplasmic in the absence of ligand, nuclear after ligand-binding and Nucleus. Localized largely in the nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • GCCR antibody
    • GCR antibody
    • GCR_HUMAN antibody
    • GCRST antibody
    • glucocorticoid nuclear receptor variant 1 antibody
    • Glucocorticoid receptor antibody
    • GR antibody
    • GRL antibody
    • Grl1 antibody
    • nr3c1 antibody
    • Nuclear receptor subfamily 3 group C member 1 antibody
    • nuclear receptor subfamily 3, group C, member 1 (glucocorticoid receptor) antibody
    see all

Anti-Glucocorticoid Receptor antibody [BuGR2] - ChIP Grade images

  • Immunocytochemistry/Immunofluorescence analysis of Glucocorticoid Receptor shows staining in A549 cells. Glucocorticoid Receptor (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2768 (1:100) over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Glucocorticoid Receptor shows staining in HeLa cells. Glucocorticoid Receptor (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2768 (1:100) over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Glucocorticoid Receptor shows staining in U251 cells. Glucocorticoid Receptor (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2768 (1:100) over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Flow cytometry analysis of Glucocorticoid Receptor showing positive staining in the nucleus and cytoplasm of NIH/3T3 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2768 at 2 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
  • Immunofluorescent analysis of Glucocorticoid Receptor using Glucocorticoid Receptor Monoclonal Antibody (BuGR2) (ab2768) shows staining in U251 Cells. Glucocorticoid Receptor (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing Glucocorticoid Receptor (ab2768) at a dilution of 1:100 over night at 4 C and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.
  • Flow cytometry analysis of Glucocorticoid Receptor showing positive staining in the nucleus and cytoplasm of Jurkat cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and then incubated with ab2768 at 1 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
  • Flow cytometry analysis of Glucocorticoid Receptor showing positive staining in the nucleus and cytoplasm of Hela cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2768 at 1 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.


  • Predicted band size : 86 kDa
    Western blot of glucocorticoid receptor on mouse liver extract using ab2768. Western blot of glucocorticoid receptor on mouse liver extract using ab2768.
  • ICC/IF image of ab2768 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2768, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing Jurkat cells stained with ab2768 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2768, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2 (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Immunohistochemical analysis of rat brain tissue, staining Glucocorticoid Receptor with ab2768.

    Tissue was fixed with formalin, permeabilized with 0.3% Triton X-100 and blocked with 3% BSA for 1 hour at room temperature; antigen retrieval was via an enzymatic method. Samples were incubated with primary antibody (1/100) for 12 hours at 4°C. A biotinylated goat anti-mouse polyclonal IgG (1/200) was used as the secondary antibody.

    See Abreview

References for Anti-Glucocorticoid Receptor antibody [BuGR2] - ChIP Grade (ab2768)

This product has been referenced in:
  • Zhou L  et al. Kaiso represses the expression of glucocorticoid receptor via a methylation-dependent mechanism and attenuates the anti-apoptotic activity of glucocorticoids in breast cancer cells. BMB Rep 49:167-72 (2016). Human . Read more (PubMed: 26424557) »
  • Wang HN  et al. Repetitive transcranial magnetic stimulation ameliorates anxiety-like behavior and impaired sensorimotor gating in a rat model of post-traumatic stress disorder. PLoS One 10:e0117189 (2015). WB ; Rat . Read more (PubMed: 25659132) »

See all 11 Publications for this product

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Application Immunocytochemistry/ Immunofluorescence
Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Sample Rat Cell (Neuron)
Specification Neuron
Permeabilization Yes - 0.25% Triton-X
Fixative Paraformaldehyde
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Submitted Jan 24 2014



We have a customer review (Abreview) using this antibody in IHC-Fr which notes successful use of enzymatic retrieval in rat brain sections.

Here is a link to this customer review:
http://www.abcam.com/Glucocorticoid-Receptor-Bu...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Brain)
Specification Brain
Fixative Formaldehyde
Antigen retrieval step Enzymatic
Permeabilization Yes - Triton X-100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
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Abcam user community

Verified customer

Submitted Oct 23 2012

I am glad you have found the information provide of use.

If you do have any further questions, please do not hesitate to contact us again.

Until then, I wish you all the best with your research.

Thank you for contacting us yesterday and sorry for the delay in getting back to you.

I can now share with you the information we have in regards to the epitope recognised by the Anti-Glucocorticoid Receptor antibody [BuGR2] (ab2768).
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Je suis ravi que vous souhaitiez participer à notre offre, tester ab2768 en IHC-FoFr et partager vos résultats. Le code promotionnel correspondant est *******, il est valable 4 mois et po...

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Merci de votre intérêt pour ab2768.
Comme expliqué par téléphone, cet anticorps a été testé en IHC-FoFr (PubMed: 20307510) mais nous ne disposons malheureusement pas d'image pour cette apllication.
Si vous souhaitez tester ab2768 en IHC-FoFr, ...

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Could you please ask customer, if they are happy to trying a different lot?

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Thank you for your enquiry regarding ab2768 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

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