Key features and details
- Mouse monoclonal [AR 441] to Androgen Receptor
- Suitable for: Flow Cyt, ICC/IF, IHC-P, WB
- Reacts with: Human
- Isotype: IgG1
Product nameAnti-Androgen Receptor antibody [AR 441]
See all Androgen Receptor primary antibodies
DescriptionMouse 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 reactivityReacts with: Human
Predicted to work with: Macaque monkey
- Prostate Carcinoma.
Storage instructionsShipped 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 bufferpH: 7.50
Preservative: 0.09% Sodium azide
Concentration information loading...
PurityProtein G purified
Purification notespurified immunoglobulin fraction.
Clone numberAR 441
Light chain typeunknown
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.
|Flow Cyt||Use at an assay dependent concentration.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-P||1/25 - 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
Upper band at 100-110KD corresponds to the "wild type AR." Lower band 70-75 KDa corresponds to a "truncated AR".
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.
modificationsSumoylated 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
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.
- AIS antibody
- ANDR_HUMAN antibody
- Androgen nuclear receptor variant 2 antibody
Anti-Androgen Receptor antibody [AR 441] (ab9474) at 1/200 dilution + Human prostate cancer cell line at 15 µg
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
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 human prostate by IHC-P.
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.
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.
ab9474 has been referenced in 31 publications.
- Younas K et al. Delayed endometrial decidualisation in polycystic ovary syndrome; the role of AR-MAGEA11. J Mol Med (Berl) 97:1315-1327 (2019). PubMed: 31256208
- Cao L et al. A high AR:ERa or PDEF:ERa ratio predicts a sub-optimal response to tamoxifen therapy in ERa-positive breast cancer. Cancer Chemother Pharmacol 84:609-620 (2019). PubMed: 31297554
- Liu X et al. Androgen receptor promotes oral squamous cell carcinoma cell migration by increasing EGFR phosphorylation. Onco Targets Ther 12:4245-4252 (2019). PubMed: 31239703
- Wang R et al. Androgen Receptor Promotes Gastric Carcinogenesis via Upregulating Cell Cycle-Related Kinase Expression. J Cancer 10:4178-4188 (2019). PubMed: 31413736
- Giatromanolaki A et al. CYP17A1 and Androgen-Receptor Expression in Prostate Carcinoma Tissues and Cancer Cell Lines. Curr Urol 13:157-165 (2019). PubMed: 31933595