Anti-Glucocorticoid Receptor beta antibody (ab3581)

Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab3581 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 Use a concentration of 5 µg/ml.

Fixation: 4% paraformaldehyde for 15 minutes at room temperature. Permeabilisation: PBS containing 0.01% saponin for 15 mins Blocking: 1% BSA in PBS Primary: overnight at 4ºC

IHC-P Use at an assay dependent concentration. PubMed: 20438742Sample Preparation: endogenous peroxidase blocked with 0.3% hydrogen peroxide. Antigen retrieval: Heat induced antigen retrieval performed in 0.01M Tris-EDTA pH 9.0. Primary antibody: overnight at 4°C
Flow Cyt Use at an assay dependent concentration.
Gel supershift assays Use at an assay dependent concentration.
ICC 1/1500.
IP Use at an assay dependent concentration. Sample preparation: RIPA buffer containing protease inhibitors was added to cell lysate. Primary antibody: 1:10 dilution for 2 h on ice Prewashed magnetic beads added to lysate mixture and incubated overnight at +4ºC. After washing, sample reducing buffer was added and boiled (5 min, 95ºC)
WB 1/500. Detects a band of approximately 90 kDa (predicted molecular weight: 83 kDa).Can be blocked with Human Glucocorticoid Receptor beta peptide (ab39765). Sample preparation: Lysates boiled in reducing buffer for 5 min. 20µg of protein loaded Blocking agent: 5% non-fat milk in Tris buffered saline (TBS) Primary antibody: overnight at 4°C

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
    • glucocorticoid nuclear receptor variant 1 antibody
    • Glucocorticoid receptor antibody
    • Glucocorticoid receptor beta isoform antibody
    • GR antibody
    • GRL antibody
    • nr3c1 antibody
    • Nuclear receptor subfamily 3 group C member 1 antibody
    see all

Anti-Glucocorticoid Receptor beta antibody images

  • ICC/IF image of ab3581 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab3581, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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.

  • Anti-Glucocorticoid Receptor beta antibody (ab3581) at 1/500 dilution + COS-74 whole cell extract transfected with Glucocorticoid Receptor beta

    Predicted band size : 83 kDa

References for Anti-Glucocorticoid Receptor beta antibody (ab3581)

This product has been referenced in:
  • Butler CA  et al. Glucocorticoid receptor ß and histone deacetylase 1 and 2 expression in the airways of severe asthma. Thorax 67:392-8 (2012). ICC/IF ; Human . Read more (PubMed: 22156779) »
  • Melarangi T  et al. Glucocorticoid resistance in chronic lymphocytic leukaemia is associated with a failure of upregulated Bim/Bcl-2 complexes to activate Bax and Bak. Cell Death Dis 3:e372 (2012). WB ; Human . Read more (PubMed: 22898870) »

See all 4 Publications for this product

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