Overview

  • Product name
    Anti-Glucocorticoid Receptor antibody - ChIP Grade
    See all Glucocorticoid Receptor primary antibodies
  • Description
    Rabbit polyclonal to Glucocorticoid Receptor - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ChIP, Flow Cyt, ICC, IP, WB, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Non human primates
    Predicted to work with: Rabbit, Guinea pig
  • Immunogen

    Synthetic peptide corresponding to Human Glucocorticoid Receptor aa 245-259.
    Sequence:

    CKPLILPDTKPKIKD


    (Peptide available as ab5833)

Properties

Applications

Our Abpromise guarantee covers the use of ab3579 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
EMSA Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration. See Lerner et al., 2003.
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC Use at an assay dependent concentration.
IP Use a concentration of 5 µg/ml.
WB Use a concentration of 5 µg/ml. Predicted molecular weight: 97 kDa.Can be blocked with Glucocorticoid Receptor peptide (ab5833). Detects an ~97 kDa protein representing GR as well as two other unidentified proteins at ~135 to ~145 kDa from rat liver extract. These cross reacting proteins are not believed to be GR or GR precursors as they do not bind [3H]dexamethasone mesylate.
IHC-Fr Use a concentration of 1 µg/ml. Staining of GR in rat hippocampus results in staining primarily in the nucleus.
ICC/IF 1/40. PubMed: 19202080

Target

  • Function
    Receptor 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 specificity
    Widely 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 disease
    Defects 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 similarities
    Belongs to the nuclear hormone receptor family. NR3 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • Domain
    Composed 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 localization
    Cytoplasm. 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

Images

  • All lanes : Anti-Glucocorticoid Receptor antibody - ChIP Grade (ab3579) at 1 µg/ml

    Lane 1 : T47D cell lysate with 5% Milk in TBST
    Lane 2 : A594 cell lysate with 5% Milk in TBST
    Lane 3 : K562 cell lysate with 5% Milk in TBST
    Lane 4 : MCF7 cell lysate with 5% Milk in TBST
    Lane 5 : Jurkat cell lysate with 5% Milk in TBST

    Secondary
    All lanes : HRP conjugate at 1/1000 dilution

    Predicted band size: 97 kDa

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labeling Glucocorticoid Receptor (green) with ab3579 at 1/100. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control. 

  • Immunocytochemistry/Immunofluorescence analysis of A2058 cells labeling Glucocorticoid Receptor (green) with ab3579 at 1/20. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control. 

References

This product has been referenced in:
  • Tobin SW  et al. Regulation of Hspb7 by MEF2 and AP-1: implications for Hspb7 in muscle atrophy. J Cell Sci 129:4076-4090 (2016). Read more (PubMed: 27632998) »
  • Li Y  et al. IL-7 Receptor Mutations and Steroid Resistance in Pediatric T cell Acute Lymphoblastic Leukemia: A Genome Sequencing Study. PLoS Med 13:e1002200 (2016). WB ; Human . Read more (PubMed: 27997540) »
See all 21 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Question
Answer

Thank you foryour cooperation.

I am sorry ab3578 did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with one unit of ab3579. The order number ****.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

Thank you for your enquiry and your interest in our products.
This antibody has not been characterized for ChIP in our lab. The ChIP application has been added to the datasheet based on this specific article:
http://www.ncbi.nlm.nih.gov/pubmed?term=14522952
There is another recent publication about this product; your customer may wish to take a look at it:
O'Brien KB et al. CARM1 is required for proper control of proliferation and differentiation of pulmonary epithelial cells. Development 137:2147-56 (2010). ChIP; Mouse. PubMed: 20530543
I hope this helps and if I can assist further, please do not hesitate to contact me.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - nuclear (LNCaP-1F5 Cell line)
Specification
LNCaP-1F5 Cell line
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Detection step
Real-time PCR
Positive control
PSA enhancer ~4 kb upstream of TSS
Negative control
Negative control region on Chromosome X not known to be bound by any nuclear receptors.

Abcam user community

Verified customer

Submitted Feb 02 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (LNCaP-1F5 Cell line and A549)
Loading amount
40 µg
Specification
LNCaP-1F5 Cell line and A549
Treatment
Ethanol, 100 nM Dexamethasone, in LNCaP-1F5 and A549 - 24 hours
Gel Running Conditions
Reduced Denaturing (7.5%)
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: +4°C

Dr. Biswajyoti Sahu

Verified customer

Submitted Nov 05 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (Neurons)
Loading amount
20 µg
Specification
Neurons
Gel Running Conditions
Reduced Denaturing (8% PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 26 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Mouse Tissue lysate - whole (skin epidermis)
Total protein in input
500 µg
Specification
skin epidermis
Immuno-precipitation step
Protein A/G

Abcam user community

Verified customer

Submitted Oct 06 2008

Answer

Thank you for your interests in our products. Yes, we have a product (ab3580) that specifically detects GR-alpha from human, mouse and rat tissues and does not detect GR-beta isoform. I have attached a datasheet link to this product for your easy access. As you know, the human GR exists in two forms, alpha and beta, which are thought to be the result of alternative splicing of a single gene. Sequence analysis indicates that the alpha and beta forms of human GR are 777 and 742 amino acids long, respectively. They are identical up to residue 727, after which they diverge. After ligand binding, the 94 kDa GR alpha isoform has been observed to translocate from the cytoplasm to the nucleus where it regulates gene expression. In contrast, the 90 kDa GR beta isoform does not appear to bind either glucocorticoid agonists or antagonists, and has been localized predominantly in the nucleus independent of hormone treatment in some human cell lines. Studies suggest that human GR beta might function as a dominant negative inhibitor of GR alpha activity.

Read More

Question
Answer

Thank you for your patience. I have received a reply from the originator of ab3579 ((Glucocorticoid Receptor Antibody)). At this time, we do not have any data regarding the affinity constant of this antibody. I apologize for any inconvenience that this may cause you. Please don't hesitate to contact us again if you have any further questions.

Read More

Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. Rabbit polyclonal to Glucocorticoid Receptor (ab3579) is a good selling antibody. It is yet to be tested by IHC using paraffin embedded sections but has been shown to work using frozen sections. I find the fact that you are observing a distinct focus strange given that this is a cytoplasmic protein that becomes nuclear following the binding of its ligand. I would like to recommend that you fix your rat brain tissue in a mild fixative such as a 5 minute incubation of ice cold acetone or alternatively methanol. This approach does not carry the potential epitope modification caused by paraformaldehyde or formalin and does not require antigen retrieval. Can you please confirm that you have been using a secondary antibody against rabbit IgG as this was not detailed in your questionaire. Furthermore you have not provided details of the dilution. I would recommend a dilution series encompassing 1/100 to 1/1000 I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More

1-10 of 13 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up