Key features and details
- Mouse monoclonal [156C135.2] to Androgen Receptor (phospho S210 + S213)
- Suitable for: WB, IHC-P
- Reacts with: Human
- Isotype: IgG
Product nameAnti-Androgen Receptor (phospho S210 + S213) antibody [156C135.2]
See all Androgen Receptor primary antibodies
DescriptionMouse monoclonal [156C135.2] to Androgen Receptor (phospho S210 + S213)
Tested applicationsSuitable for: WB, IHC-Pmore details
Species reactivityReacts with: Human
Predicted to work with: Rat, Rabbit, Horse, Guinea pig, Hamster, Cat, Dog, Pig, Chimpanzee, Monkey, Baboon, Cynomolgus monkey, Hedgehog, Rhesus monkey
- IGF1 stimulated LNCaP cells. IHC-P: human normal prostate FFPE tissue sections
In IGF-1 stimulated LNCaP cells (passage number 38), a ~110 kDa band was observed. The serine phosphorylation site recognized by ab45089 has been alternatively referred to Ser213 (Lee and Chang, 2003) and Ser210 (Lin et al, 2003). Variations in denotation can arise from how the sequence is counted in various GenBank accession numbers. The site is denoted as Ser213 in GenBank Accession No. A39248, which was used to design the immunogen.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
Storage bufferPreservative: 0.05% Sodium azide
Constituents: PBS, 0.05% BSA
Concentration information loading...
PurityProtein G purified
Primary antibody notesIn IGF-1 stimulated LNCaP cells (passage number 38), a ~110 kDa band was observed. The serine phosphorylation site recognized by ab45089 has been alternatively referred to Ser213 (Lee and Chang, 2003) and Ser210 (Lin et al, 2003). Variations in denotation can arise from how the sequence is counted in various GenBank accession numbers. The site is denoted as Ser213 in GenBank Accession No. A39248, which was used to design the immunogen.
Our Abpromise guarantee covers the use of ab45089 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 - 4 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 99 kDa).|
|IHC-P||Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
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.
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All lanes : Anti-Androgen Receptor (phospho S210 + S213) antibody [156C135.2] (ab45089) at 1 µg/ml
Lane 1 : LNCaP cells with serum starved cells
Lane 2 : LNCaP cells with IGF1 incubated for 4h at 0.1 µg/ml
Lane 3 : LNCaP cells with incubated with 20 ug LY294002 for 30 min then IGF1 incubated for 4h at 0.1 µg/ml
Predicted band size: 99 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
IHC image of Androgen Receptor (phospho S213 + S210) staining in human normal prostate formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab45089, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab45089 has been referenced in 6 publications.
- Robitaille CN et al. Antiandrogenic mechanisms of pesticides in human LNCaP prostate and H295R adrenocortical carcinoma cells. Toxicol Sci 143:126-35 (2015). PubMed: 25324206
- Hong H et al. Testosterone regulates cell proliferation in aggressive fibromatosis (desmoid tumour). Br J Cancer 104:1452-8 (2011). PubMed: 21468052
- Agapova OA et al. A neuroactive steroid 5alpha-androstane-3alpha,17beta-diol regulates androgen receptor level in astrocytes. J Neurochem 98:355-63 (2006). PubMed: 16638015
- Kirkegaard T et al. Observer variation in immunohistochemical analysis of protein expression, time for a change? Histopathology 48:787-94 (2006). PubMed: 16722926
- Bhuiyan MM et al. Down-regulation of androgen receptor by 3,3'-diindolylmethane contributes to inhibition of cell proliferation and induction of apoptosis in both hormone-sensitive LNCaP and insensitive C4-2B prostate cancer cells. Cancer Res 66:10064-72 (2006). PubMed: 17047070
- Lin HK et al. Suppression versus induction of androgen receptor functions by the phosphatidylinositol 3-kinase/Akt pathway in prostate cancer LNCaP cells with different passage numbers. J Biol Chem 278:50902-7 (2003). PubMed: 14555644