Recombinant
RabMAb

Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Overview

  • Product name
    Anti-Androgen Receptor antibody [EPR1535(2)]
    See all Androgen Receptor primary antibodies
  • Description
    Rabbit monoclonal [EPR1535(2)] to Androgen Receptor
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Androgen Receptor aa 1-100 (internal sequence). The exact sequence is proprietary.
    Database link: P10275
    (Peptide available as ab191380)

  • Positive control
    • T47-D and LnCaP cell lysates human prostate tissue human prostatic adenocarcinoma tissue Rat prostate and mouse prostate lysates Rat and Mouse testis MCF7
  • 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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133273 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 1/2000. Predicted molecular weight: 98 kDa.Can be blocked with Androgen Receptor peptide (ab191380).

For unpurified use at 1:1000

IHC-P 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100-1/250

ICC/IF 1/100 - 1/250.

Target

  • Function
    Steroid hormone receptors are ligand-activated transcription factors that regulate eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Transcription factor activity is modulated by bound coactivator and corepressor proteins. Transcription activation is down-regulated by NR0B2. Activated, but not phosphorylated, by HIPK3 and ZIPK/DAPK3.
    Isoform 3 and isoform 4 lack the C-terminal ligand-binding domain and may therefore constitutively activate the transcription of a specific set of genes independently of steroid hormones.
  • Tissue specificity
    Isoform 2 is mainly expressed in heart and skeletal muscle (PubMed:15634333). Isoform 3 is expressed by basal and stromal cells of prostate (at protein level) (PubMed:19244107).
  • Involvement in disease
    Androgen insensitivity syndrome
    Spinal and bulbar muscular atrophy X-linked 1
    Defects in AR may play a role in metastatic prostate cancer. The mutated receptor stimulates prostate growth and metastases development despite of androgen ablation. This treatment can reduce primary and metastatic lesions probably by inducing apoptosis of tumor cells when they express the wild-type receptor.
    Androgen insensitivity, partial
  • 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. In the presence of bound steroid the ligand-binding domain interacts with the N-terminal modulating domain, and thereby activates AR transcription factor activity. Agonist binding is required for dimerization and binding to target DNA. The transcription factor activity of the complex formed by ligand-activated AR and DNA is modulated by interactions with coactivator and corepressor proteins. Interaction with RANBP9 is mediated by both the N-terminal domain and the DNA-binding domain. Interaction with EFCAB6/DJBP is mediated by the DNA-binding domain.
  • Post-translational
    modifications
    Sumoylated on Lys-388 (major) and Lys-521. Ubiquitinated. Deubiquitinated by USP26. 'Lys-6' and 'Lys-27'-linked polyubiquitination by RNF6 modulates AR transcriptional activity and specificity.
    Phosphorylated in prostate cancer cells in response to several growth factors including EGF. Phosphorylation is induced by c-Src kinase (CSK). Tyr-535 is one of the major phosphorylation sites and an increase in phosphorylation and Src kinase activity is associated with prostate cancer progression. Phosphorylation by TNK2 enhances the DNA-binding and transcriptional activity and may be responsible for androgen-independent progression of prostate cancer. Phosphorylation at Ser-83 by CDK9 regulates AR promoter selectivity and cell growth. Phosphorylation by PAK6 leads to AR-mediated transcription inhibition.
    Palmitoylated by ZDHHC7 and ZDHHC21. Palmitoylation is required for plasma membrane targeting and for rapid intracellular signaling via ERK and AKT kinases and cAMP generation.
  • Cellular localization
    Nucleus. Cytoplasm. Predominantly cytoplasmic in unligated form but translocates to the nucleus upon ligand-binding. Can also translocate to the nucleus in unligated form in the presence of RACK1.
  • Information by UniProt
  • Database links
  • Form
    There are 2 isoforms produced by alternative splicing. Isoform 1 is also known as: AR-B; isoform 2 is known as AR-A or variant AR45.
  • Alternative names
    • AIS antibody
    • ANDR_HUMAN antibody
    • Androgen nuclear receptor variant 2 antibody
    • Androgen receptor (dihydrotestosterone receptor; testicular feminization; spinal and bulbar muscular atrophy; Kennedy disease) antibody
    • Androgen receptor antibody
    • androgen receptor splice variant 4b antibody
    • AR antibody
    • AR8 antibody
    • DHTR antibody
    • Dihydro testosterone receptor antibody
    • Dihydrotestosterone receptor (DHTR) antibody
    • Dihydrotestosterone receptor antibody
    • HUMARA antibody
    • HYSP1 antibody
    • KD antibody
    • Kennedy disease (KD) antibody
    • NR3C4 antibody
    • Nuclear receptor subfamily 3 group C member 4 (NR3C4) antibody
    • Nuclear receptor subfamily 3 group C member 4 antibody
    • SBMA antibody
    • SMAX1 antibody
    • Spinal and bulbar muscular atrophy (SBMA) antibody
    • Spinal and bulbar muscular atrophy antibody
    • Testicular Feminization (TFM) antibody
    • TFM antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Androgen receptor with purified ab133273 at 1:100 dilution (1.3μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/5000 dilution (purified) + Mouse prostate lysates at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 98 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded human prostatic adenocarcinoma tissue labelling Androgen Receptor  using  unpurified ab133273 at 1/100 dilution

  • Immunofluorescent staining of LnCAP cells labelling Androgen Receptor using unpurified ab133273, at 1/100 dilution

  • All lanes : Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/5000 dilution (purified)

    Lane 1 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates
    Lane 2 : Rat prostate lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size: 98 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue sections labeling Pumilio 1 with Purified ab133273 at 1:500 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue sections labeling Pumilio 1 with Purified ab133273 at 1:500 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostaitc hyperplasia tissue sections labeling Pumilio 1 with Purified ab133273 at 1:500 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/1000 dilution (unpurified) + LNCaP (Human prostate carcinoma epithelial cell) at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 98 kDa
    Observed band size: 110 kDa (why is the actual band size different from the predicted?)


    Exposure time: 3 seconds


    Blocking/Diluting buffer 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded human prostate tissue labelling Androgen Receptor using unpurified ab133273, at1/100 dilution.

  • Unpurified  ab133273 showing positive staining in Breast carcinoma tissue.

  • Unpurified  ab133273 showing negative staining in Normal brain tissue.

  • Unpurified  ab133273 showing negative staining in Normal tonsil tissue.

  • Unpurified ab133273 showing positive staining in Endometrial carcinoma tissue.

  • Unpurified ab133273 showing negative staining in Normal breast tissue.

  • Unpurified ab133273 showing negative staining in Normal colon tissue.

  • Unpurified ab133273 showing negative staining in Ovarian carcinoma tissue.

  • Unpurified ab133273 showing negative staining in Colonic adenocarcinoma tissue.

  •  Unpurified ab133273 showing negative staining in Normal liver tissue.

  • Unpurified  ab133273 showing positive staining in Prostatic carcinoma T3 tissue.

References

This product has been referenced in:
  • Patel D  et al. The histone demethylase LSD1 regulates inner ear progenitor differentiation through interactions with Pax2 and the NuRD repressor complex. PLoS One 13:e0191689 (2018). Read more (PubMed: 29370269) »
  • Shao L  et al. Fibroblast growth factor receptor signaling plays a key role in transformation induced by the TMPRSS2/ERG fusion gene and decreased PTEN. Oncotarget 9:14456-14471 (2018). Read more (PubMed: 29581856) »

See all 16 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cow Tissue sections (Sertoli cells in testis)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate buffer pH 6.0
Permeabilization
No
Specification
Sertoli cells in testis
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative
Bouins solution
Username

Natasja Costermans

Verified customer

Submitted Sep 14 2016

Thank you very much for your interest our products.


To our knowledge, ab133273 not been tested in IHC-FrFl (Immunohistochemistry - Free Floating).

However by participating in our AbTrial program you can now use our products in...

Read More

Thank you for your inquiry.

I can confirm that in our hands ab52615 (1852-1) is not working in IHC, ICC and IP. Therefore this antibody is not covered by our guarantee since we are not convinced that ab52615 is suitable for these applications...

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

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