Overview

  • Product name
    Anti-Glucocorticoid Receptor antibody
    See all Glucocorticoid Receptor primary antibodies
  • Description
    Rabbit polyclonal to Glucocorticoid Receptor
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IHC-P, Inhibition Assay, ChIP, IP, ICC, WB, IHC-FoFrmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Rabbit, Hamster, Cow, Human, Saccharomyces cerevisiae, Non human primates
    Predicted to work with: Guinea pig, Pig
  • Immunogen

    Synthetic peptide corresponding to Human Glucocorticoid Receptor aa 346-367.
    Sequence:

    DQKPIFNVIPPIPVGSENWNRC


    (Peptide available as ab5019)

Properties

Applications

Our Abpromise guarantee covers the use of ab3578 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 1/20.
IHC-P 1/200.
Inhibition Assay Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ICC Use a concentration of 1 µg/ml.

Immunocytochemical staining of GR in HeLa cells results in cytoplasmic staining in the absence of ligand, and nuclear staining after hormone administration.

WB Use a concentration of 5 µg/ml. Detects a band of approximately 95 kDa (predicted molecular weight: 86 kDa).Can be blocked with Glucocorticoid Receptor peptide (ab5019).
EMSA Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration. PubMed: 18279320

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

  • Immunocytochemistry/Immunofluorescence analysis of U251 cells labeling Glucocorticoid (green) with ab3578 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.

  • Immunocytochemistry/Immunofluorescence analysis of NIH-3T3 cells labeling Glucocorticoid (green) with ab3578 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.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labeling Glucocorticoid (green) with ab3578 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.

  • ab3578 staining Glucocorticoids Receptors in Mouse brain tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue underwent fixation in 4% paraformaldehyde and 15µm sections were cut using cryostat. The primary antibody was diluted, 1/100 in TNB buffer and incubated with sample for 18 hours at 4°C. A HRP conjugated goat polyclonal to rabbit IgG, diluted 1/500 was used as secondary. Red staining repersents staining with ab3578 in perilesional cells. 

    See Abreview

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human cervical carcinoma tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing Glucocorticoid Receptor (ab3578) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing Glucocorticoid Receptor (ab3578) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

References

This product has been referenced in:
  • Dayger CA  et al. Physiological correlates of reproductive decisions: Relationships among body condition, reproductive status, and the hypothalamus-pituitary-adrenal axis in a reptile. Horm Behav 100:1-11 (2018). WB . Read more (PubMed: 29452075) »
  • Zhao M  et al. Iron-induced neuronal damage in a rat model of post-traumatic stress disorder. Neuroscience 330:90-9 (2016). WB, IHC ; Rat . Read more (PubMed: 27208615) »
See all 8 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Western blot
Sample
Red sided garter snake Tissue lysate - whole (heart, brain, liver)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
5 µg
Specification
heart, brain, liver
Blocking step
5% milk + 1% BSA in Tris-buffered saline pH 7.4 as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1µg/mL · Temperature: 24°C

Abcam user community

Verified customer

Submitted Dec 06 2016

Question
Answer

Thank you for your enquiry and interest in our products.
The immunogen sequence used to raise this antibody (ab3578: Anti-Glucocorticoid Receptor) was DQKPIFNVIPPIPVGSENWNR, corresponding to amino acids 346-367 of Human Glucocorticoid Receptor. However, currently this peptide is not in our catalogue.
Apologies!
If you need any further assistance in the future, please do not hesitate to contact me.

Read More

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 taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

I apologise for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation.

Please note that the only lot we currently have in stock for ab3578 is the same as the one you received and tested (lot # GR76455). Therefore I can offer you as a replacement an alternative anti-Glucocorticoid Receptor from the following list :



https://www.abcam.com/Glucocorticoid-Receptor-antibody-ChIP-Grade-ab3579.html , Rabbit polyclonal




https://www.abcam.com/Glucocorticoid-Receptor-phospho-S226-antibody-ab53692.html https://www.abcam.com/Glucocorticoid-Receptor-phospho-S226-antibody-ab53692.html Rabbit polyclonal




https://www.abcam.com/Glucocorticoid-Receptor-antibody-ab16510.html, Rabbit polyclonal


, Rabbit polyclonal

, Rabbit monoclonal
https://www.abcam.com/Glucocorticoid-Receptor-antibody-BuGR2-ab2768.html, Mouse monoclonal



https://www.abcam.com/Glucocorticoid-Receptor-antibody-ab55400.html
https://www.abcam.com/Glucocorticoid-Receptor-antibody-EPR4595-ab109022.html



Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Question
Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WBand Mouse, Rat, Hamster and Human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you are also able to provide an image which would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionaire.



Order Details
Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

Read More

Answer

I just went back and checked, when I thought I was replying to you, I was actually replying to the customer. Sorry about that. I have processed the two credits for ab3578. If there is anything else I can help you with, please let me kn

Read More

Answer

I got the email you sent on Monday, the customer must have forwarded it to me. Please see below for what I replied with on Monday: "Thank you for your reply. Before I issue the credit, I just want to confirm that we are talking about the correct antibody. I thought we were talking about ab3578, however in the last email you mentioned ab3574. I am sure it was just a typo, but I wanted to double check. Also could you provide me with the customers Cedarlane number, so that I can add that information to the credit note for ease of tracking. I look forward to your reply." If you could just clarify the issue above, I can go ahead and process the credit.  

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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
Mouse Tissue sections (brain)
Antigen retrieval step
None
Permeabilization
No
Specification
brain
Blocking step
Perkin-Elmer TNB Blocking Reagent as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 23 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Hamster Cell (Expression of Human Glucocorticoid Receptor in CHO)
Permeabilization
Yes - 0.1 % Triton X-100
Specification
Expression of Human Glucocorticoid Receptor in CHO
Blocking step
TBS + 3% Dried Milk + 0.1 % Triton X-100 as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 22°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 29 2008

1-10 of 12 Abreviews or Q&A

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