Name: Anti-Glucocorticoid Receptor antibody [EPR19621]
Brand: RabMAb
SKU: ab183127
Rating: 5 (2 reviews)

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Western blot - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19621] to Glucocorticoid Receptor
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 647 Carrier Free

Product name

Anti-Glucocorticoid Receptor antibody [EPR19621]
See all Glucocorticoid Receptor primary antibodies
Description

Rabbit monoclonal [EPR19621] to Glucocorticoid Receptor
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal heart and fetal kidney lysates; HeLa, A549, U-87 MG, HEK-293 and A431 whole cell lysates; HEK-293 whole cell lysate transfected with human Glucocorticoid Receptor with GFP-Myc tag. IHC-P: Human glioma and cervix carcinoma tissues; Mouse liver tissue; Rat hippocampus tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19621
Isotype

IgG
Research areas
- Neuroscience
- Processes

Alternative Versions
- Anti-Glucocorticoid Receptor antibody [EPR19621] - BSA and Azide free (ab223138)
- Alexa Fluor® 647 Anti-Glucocorticoid Receptor antibody [EPR19621] (ab237225)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human NR3C1 (Glucocorticoid Receptor) knockout HeLa cell line (ab261766)
- Human NR3C1 (Glucocorticoid Receptor) knockout A549 cell line (ab261862)
KO cell lysates
- Human NR3C1 (Glucocorticoid Receptor) knockout HeLa cell lysate (ab257009)
- Human NR3C1 (Glucocorticoid Receptor) knockout A549 cell lysate (ab261671)
Recombinant Protein
- Recombinant human Glucocorticoid Receptor protein (ab82089)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab183127 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
WB	(2)	1/2000. Detects a band of approximately 86, 83 kDa (predicted molecular weight: 86 kDa).
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/500.
Flow Cyt (Intra)		1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
WB 1/2000. Detects a band of approximately 86, 83 kDa (predicted molecular weight: 86 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/500.
Flow Cyt (Intra) 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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.
Target information above from: UniProt accession P04150 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 2908 Human
- Entrez Gene: 14815 Mouse
- Entrez Gene: 24413 Rat
- Omim: 138040 Human
- SwissProt: P04150 Human
- SwissProt: P06537 Mouse
- SwissProt: P06536 Rat
- Unigene: 122926 Human
- Unigene: 129481 Mouse
- Unigene: 90070 Rat
see all
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

Western blot - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

All lanes : Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : NR3C1 knockout HeLa cell lysate
Lane 3 : Wild-type A549 cell lysate
Lane 4 : NR3C1 knockout A549 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution

Performed under reducing conditions.

Predicted band size: 86 kDa
Observed band size: 90-100 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab183127 observed at 90-100 kDa. Red - loading control ab8245 observed at 37 kDa.

ab183127 Anti-Glucocorticoid Receptor antibody [EPR19621] was shown to specifically react with Glucocorticoid Receptor in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261766 (knockout cell lysate ab257009) was used. Wild-type and Glucocorticoid Receptor knockout samples were subjected to SDS-PAGE. ab183127 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

All lanes : Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127) at 1/2000 dilution

Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : NR3C1 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 4 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 86 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab183127 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab183127 was shown to recognize NR3C1 in wild-type A549 cells as signal was lost at the expected MW in NR3C1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NR3C1 knockout samples were subjected to SDS-PAGE. Ab183127 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/1000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

All lanes : Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127) at 1/2000 dilution

Lane 1 : Human fetal heart lysate
Lane 2 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 86 kDa
Observed band size: 83,86 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure times: Lane 1: 30 seconds; Lane 2: 15 seconds.

This antibody may recognize eight isoforms. The predicted MW are from 61KDa to 86KDa in human, respectively.
Western blot - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

All lanes : Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127) at 1/2000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : A431 (Human epidermoid carcinoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 86 kDa
Observed band size: 83,86 kDa why is the actual band size different from the predicted?

Exposure time: 10 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

This antibody may recognize eight isoforms. The predicted MW are from 61KDa to 86KDa in human, respectively.
Western blot - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

All lanes : Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127) at 1/20000 dilution

Lane 1 : Empty vector with GFP-Myc tag (vector control) transfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with human Glucocorticoid Receptor with GFP-Myc tag

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 86 kDa
Observed band size: 112 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: 0.5 second.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling Glucocorticoid Receptor with ab183127 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on tumor cells of the Human glioma is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling Glucocorticoid Receptor with ab183127 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on tumor cells of the cervix carcinoma is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Glucocorticoid Receptor with ab183127 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on hepatocytes of the mouse liver is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

Immunohistochemical analysis of paraffin-embedded Rat hippocampus tissue labeling Glucocorticoid Receptor with ab183127 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on rat hippocampus is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Glucocorticoid Receptor with ab183127 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

The results show signal translocation after dexamethasone (100 nM for 2 hours) treatment on Hela cells. PMID: 24291004.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows:
-ve control 1: ab183127 at 1/500 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Glucocorticoid Receptor with ab183127 at 1/500 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A]-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor^® 488) at 1/500 dilution was used as the secondary antibody.
Anti-Glucocorticoid Receptor antibody [EPR19621] (ab183127)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab183127? Please let us know so that we can cite the reference in this datasheet.

ab183127 has been referenced in 6 publications.

Mossakowska BJ et al. Difference in miRNA Expression in Functioning and Silent Corticotroph Pituitary Adenomas Indicates the Role of miRNA in the Regulation of Corticosteroid Receptors. Int J Mol Sci 23:N/A (2022). PubMed: 35270010
Gannon AL et al. A Novel Model Using AAV9-Cre to Knockout Adult Leydig Cell Gene Expression Reveals a Physiological Role of Glucocorticoid Receptor Signalling in Leydig Cell Function. Int J Mol Sci 23:N/A (2022). PubMed: 36499341
Choi D et al. ß-Ionone Attenuates Dexamethasone-Induced Suppression of Collagen and Hyaluronic Acid Synthesis in Human Dermal Fibroblasts. Biomolecules 11:N/A (2021). PubMed: 33919331
Qu P et al. Habenula lesions improve glucose metabolism in rats with type 2 diabetes by increasing insulin sensitivity and inhibiting gluconeogenesis. BMJ Open Diabetes Res Care 8:N/A (2020). PubMed: 32393480
Hu S et al. Prenatal caffeine exposure increases the susceptibility to non-alcoholic fatty liver disease in female offspring rats via activation of GR-C/EBPa-SIRT1 pathway. Toxicology 417:23-34 (2019). PubMed: 30776459
Hu S et al. Glucocorticoid programming mechanism for hypercholesterolemia in prenatal ethanol-exposed adult offspring rats. Toxicol Appl Pharmacol 375:46-56 (2019). PubMed: 31075344

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1-2 of 2 Abreviews or Q&A

Application

Western blot

Sample

Pig Tissue lysate - whole (Liver)

Gel Running Conditions

Reduced Denaturing (10% SDS-PAGE)

Loading amount

5 µg

Specification

Liver

Blocking step

skimmed milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Liliana Kiczak

Verified customer

Submitted Feb 23 2021

Application

Western blot

Sample

Pig Cell lysate - whole cell (Porcine brain endothelial cells)

Gel Running Conditions

Reduced Denaturing (10)

Loading amount

15 µg

Specification

Porcine brain endothelial cells

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Yu Siong Ho

Verified customer

Submitted Jul 31 2019

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (6)

Customer reviews and Q&As

Western blot abreview for Anti-Glucocorticoid Receptor antibody [EPR19621]

Western blot abreview for Anti-Glucocorticoid Receptor antibody [EPR19621]

Post-translational
modifications