Anti-Androgen Receptor antibody [AR 441] (ab9474)

Overview

  • Product nameAnti-Androgen Receptor antibody [AR 441]
    See all Androgen Receptor primary antibodies
  • Description
    Mouse monoclonal [AR 441] to Androgen Receptor
  • SpecificityThis antibody is specific to a protein of 110 kD, identified as androgen receptor. This antibody reacts with full length androgen receptor and also with the newly described A form of the receptor. This antibody does not cross react with estrogen, progesterone or glucocorticoid receptors.
  • Tested applicationsSuitable for: Flow Cyt, ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Macaque Monkey
  • Immunogen

    Synthetic peptide corresponding to Human Androgen Receptor aa 299-315.
    Sequence:

    STEDTAEYSPFKGGYTK


    Database link: P10275

  • Positive control
    • Prostate Carcinoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab9474 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
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF 1/50.
IHC-P 1/25 - 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/200.

Upper band at 100-110KD corresponds to the "wild type AR." Lower band 70-75 KDa corresponds to a "truncated AR".

Target

  • FunctionSteroid 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 specificityIsoform 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 diseaseAndrogen 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 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. 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 localizationNucleus. 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
  • FormThere 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

Anti-Androgen Receptor antibody [AR 441] images

  • ab9474 staining human prostate by IHC-P.

  • Anti-Androgen Receptor antibody [AR 441] (ab9474) at 1/200 dilution + Human prostate cancer cell line at 15 µg

    Secondary
    Donkey anti-mouse IgG (H+L) at 1/200 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 100-110 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 70-75 kDa (possible truncated form).

    Exposure time : 10 minutes

    This image is courtesy of an abreview submitted by Dr. Víctor Campa.

    Blocking agent: 3% BSA, Dilution Buffer: TBST+3%BSA.

    Upper band at 100-110KD corresponds to the wild type Androgen Receptor.  Lower band 70-75 KDa corresponds to a truncated Androgen Receptor.

  •  ab9474 staining the Androgen Receptor in the human 22Rv1 Prostate Cancer Cell Line by ICC/IF (immunocytochemistry/immunofluorescence). Cells were formaldehyde fixed, permeabilized by 0.1% Triton for 15 minutes, and blocked with glycine (7.5mg/ml) for 15 minutes at room temperature. The sample was incubated with the primary antibody (1/50 in PBS) for 12 hours at 4°C. An undiluted Alexa Fluor 488®-conjugated Donkey anti-mouse polyclonal (ab150105) was used as the secondary.

    See Abreview

  • Overlay histogram showing MCF-7 cells stained with ab9474 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab9474, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was DyLight ® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Overlay histogram showing PC3 cells stained with ab9474 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab9474, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was DyLight ® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in PC3 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

References for Anti-Androgen Receptor antibody [AR 441] (ab9474)

This product has been referenced in:
  • Olsen JR  et al. Context dependent regulatory patterns of the androgen receptor and androgen receptor target genes. BMC Cancer 16:377 (2016). WB, ICC/IF ; Human . Read more (PubMed: 27378372) »
  • Fu Y  et al. Ductal activation of oncogenic KRAS alone induces sarcomatoid phenotype. Sci Rep 5:13347 (2015). Read more (PubMed: 26289340) »

See all 10 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (DU145 Prostate cancer cell line)
Permeabilization Yes - 0.1% v/v TritonX-100
Specification DU145 Prostate cancer cell line
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative Paraformaldehyde
Username

Dr. Dimitra Kalamida

Verified customer

Submitted May 30 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Sample Human Cultured Cells (prostate cancer cell line(LNCaP))
Specification prostate cancer cell line(LNCaP)
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 23 2013

There are two options for staining a tissue section with two mouse monoclonal antibodies. The first is to directly conjugate the antibodies, removing the need for any secondary antibodies. We have conjugation kits that go by the name "EasyLink". Here i...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (C4-2B (Prostate cancer cell line))
Specification C4-2B (Prostate cancer cell line)
Fixative Paraformaldehyde
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jun 18 2012

Thank you for your inquiry.

We recommend to use this antibody at a dilution of 1/50in IHC-P. This is only a recommended starting dilution and may well have to be optimized by your customer according to their needs and samples.

If th...

Read More

Thank you for your interest. It is very likely that ab9474 works in mouse but we have no data yet. According to Blast search, the homology between human and mouse AR is very high. http://www.uniprot.org/blast/uniprot/2011101050RXMELAA8 ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Prostate carcinoma)
Specification Prostate carcinoma
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer pH6.0
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Dec 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Prostate Cancer cell line)
Loading amount 15 µg
Specification Prostate Cancer cell line
Gel Running Conditions Reduced Denaturing (7%)
Blocking step BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Dr. Víctor Campa

Verified customer

Submitted Dec 30 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (22Rv1 Prostate Cancer Cell line)
Specification 22Rv1 Prostate Cancer Cell line
Fixative Formaldehyde
Permeabilization Yes - 0.1%Triton 15 minutes
Blocking step (agent) for 15 minute(s) · Concentration: 7500µg/mL · Temperature: RT°C
Username

Dr. Víctor Campa

Verified customer

Submitted Dec 02 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (C4-2b (prostate cancer cell line))
Specification C4-2b (prostate cancer cell line)
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% triton
Blocking step BSA as blocking agent for 15 minute(s) · Concentration: 0.2% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jun 24 2008

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