Recombinant
RabMAb

Recombinant Anti-Glucocorticoid Receptor (phospho S226) antibody [EPR22216-414] (ab228972)

Overview

  • Product name

    Anti-Glucocorticoid Receptor (phospho S226) antibody [EPR22216-414]
    See all Glucocorticoid Receptor primary antibodies
  • Description

    Rabbit monoclonal [EPR22216-414] to Glucocorticoid Receptor (phospho S226)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Dot blotmore details
    Unsuitable for: ChIP,Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Glucocorticoid Receptor aa 200-300. The exact sequence is proprietary.
    Database link: P04150

  • Positive control

    • WB: U-2 OS treated with 1 µM Dexamethasone for 1hour whole cell lysate 20 µg (Untreated membrane), cell lysate. Dot blot: Glucocorticoid Receptor (phospho S226) peptide (aa220-231) and Glucocorticoid Receptor (phospho S226) peptide (aa225-234). IHC-P: Human endometrial cancer tissue.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab228972 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 1/1000. Predicted molecular weight: 85 kDa.

 

 

IHC-P 1/1000.

 

 

Dot blot 1/1000.

 

 

  • Application notes
    Is unsuitable for ChIP,Flow Cyt,ICC/IF or IP.
  • 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 (phospho S226) antibody [EPR22216-414] (ab228972) at 1/1000 dilution

      Lane 1 : Untreated U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate (Untreated membrane)
      Lane 2 : U-2 OS treated with 1 µM Dexamethasone for 1hour whole cell lysate 20 µg (Untreated membrane)
      Lane 3 : U-2 OS treated with 1 µM Dexamethasone for 1hour whole cell lysate 20 µg (Phosphatase treated membrane

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



      Blocking and diluting buffer and concentration: 5% NFDM/TBST

      Glucocorticoid receptor is phosphorylated upon stimulation of dexamethasone, a corticosteroid (PMID: 28781762).  Exposure time: 3 minutes

    • Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labeling Glucocorticoid Receptor (phospho S226) with ab228972 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human endometrium cancer (panel A), and no staining after alkaline phosphatase treatment (panel B) (PMID:12351702, 12225995). The section was incubated with ab228972 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    • Dot blot analysis using ab228972 at 1/1000 dilution. The secondary antibody was a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.

      Lane 1: Glucocorticoid Receptor (phospho S226) peptide (aa220-231)
      Lane 2: Glucocorticoid Receptor (phospho S226) peptide (aa225-234)
      Lane 3: Glucocorticoid Receptor non-phospho peptide (aa220-234)

      Blocking/diluting buffer and concentration: 5% NFDM/TBST
      Exposure time: 3 minutes.

    References

    ab228972 has not yet been referenced specifically in any publications.

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