Name: Anti-Androgen Receptor antibody [EPR1535(2)]
Brand: RabMAb
SKU: ab133273
Rating: 4 (1 reviews)

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 (N terminal). The exact sequence is proprietary.
Database link: P10275
(Peptide available as ab191380)
Positive control
- WB: T47-D, LnCaP and 22Rv1 cell lysates; Rat and mouse prostate lysates. IHC-P: Human prostate, prostatic adenocarcinoma, prostatic hyperplasia tissues; Rat and mouse testis tissues; Breast carcinoma and prostatic carcinoma T3 tissues. ICC/IF: MCF7 cells;EP156T-AR, 957E/hTERT-AR and LNCaP cells.
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.

Form

Liquid
Storage instructions

Shipped 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 buffer

pH: 7.40
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR1535(2)
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Immunizing Peptide (Blocking)
- Androgen Receptor peptide (ab191380)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- LNCaP nuclear extract lysate (ab14857)
- T47D nuclear extract lysate (ab14896)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

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

1/2000. Predicted molecular weight: 98 kDa.Can be blocked with Androgen Receptor peptide (ab191380).

For unpurified use at 1:1000

For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).

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.

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.
Target information above from: UniProt accession P10275 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 367 Human
- Entrez Gene: 11835 Mouse
- Entrez Gene: 24208 Rat
- Omim: 313700 Human
- SwissProt: P10275 Human
- SwissProt: P19091 Mouse
- SwissProt: P15207 Rat
- Unigene: 76704 Human
- Unigene: 39005 Mouse
- Unigene: 394224 Mouse
- Unigene: 439657 Mouse
- Unigene: 9813 Rat
see all
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

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

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
Immunocytochemistry/ Immunofluorescence - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)Image from Azeem W et al., PLoS One. 2017;12(6):e0177861. Fig 6.; doi: 10.1371/journal.pone.0177861.

Texas red (Tx-Red) indirect immunofluorescent detection of Androgen Receptor using ab133273 in (A) EP156T-AR, (B) 957E/hTERT-AR and (C) LNCaP cells. The cells were treated with ± 1 nM R1881 for 24 hours.
Immunocytochemistry/ Immunofluorescence - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Immunohistochemical analysis of paraffin-embedded human prostatic adenocarcinoma tissue labelling Androgen Receptor using unpurified ab133273 at 1/100 dilution
Western blot - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

All lanes : Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/20000 dilution

Lane 1 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method
Lane 2 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method
Lane 3 : 22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method
Lane 4 : 22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method

Lysates/proteins at 1/15 dilution per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 98 kDa
Observed band size: 120 kDa
why is the actual band size different from the predicted?

Exposure time: 10 seconds

Blocking/Diluting buffer - 5% NFDM/TBST

The androgen receptor variant band detected in 22RV1 cells is reported by PMID: 22315407.

We recommend you to try both RIPA and 1%SDS Hot lysis preparation methods to get desired bands.
Immunocytochemistry/ Immunofluorescence - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Immunofluorescent staining of LnCAP cells labelling Androgen Receptor using unpurified ab133273, at 1/100 dilution
Western blot - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

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

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

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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic hyperplasia tissue sections labeling Androgen Receptor 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.
Western blot - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Immunohistochemical analysis of paraffin-embedded human prostate tissue labelling Androgen Receptor using unpurified ab133273, at1/100 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Unpurified ab133273 showing positive staining in Breast carcinoma tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Unpurified ab133273 showing negative staining in Normal brain tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Unpurified ab133273 showing negative staining in Normal tonsil tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Unpurified ab133273 showing positive staining in Endometrial carcinoma tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Unpurified ab133273 showing negative staining in Normal breast tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Unpurified ab133273 showing negative staining in Normal colon tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Unpurified ab133273 showing negative staining in Ovarian carcinoma tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Unpurified ab133273 showing negative staining in Colonic adenocarcinoma tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Unpurified ab133273 showing negative staining in Normal liver tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)

Unpurified ab133273 showing positive staining in Prostatic carcinoma T3 tissue.

Western blot protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Protocol Booklet

Datasheet

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). IHC . Read more (PubMed: 29581856) »

See all 23 Publications for this product

Publishing research using ab133273? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

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

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

Ms. Natasja Costermans

Verified customer

Submitted Sep 14 2016

Question

Thank you very much for your reply.

I would like to use the antibodies against Androgen receptor for detection of androgen receptor in mouse brain floating tissue sections. Is you warranty covering also this application? On your website you mostly show images for prostate, which has much higher expression levels than the brain, so therefore, I would like to check whether your warranty also cover brain tissue for that specific antibody.

Interestingly, though, some our colleagues did use antibody 1852-1 successfully (at least they showed photomicrographs) for detecting AR in mouse brain tissue.

Best regards,

Abcam community

Verified customer

Asked on Nov 05 2012

Answer

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 an untested application or species without financial risk.

Simply follow these easy steps below to apply for our AbTrial Program:

1. Reply to this email, letting us know you are interested in testing this product.

2. Our scientists will email you an inactive personal discount code for the value of the product.

3. Purchase and test the product at the regular price.

4. Submit your results, including your discount code in the additional notes section of your Abreview.

5. Once the Abreview is submitted, the discount code will become active.

6. Apply your discount code on your next order to receive that value off.

Please let me know if you have any questions about this offer and I would be happy to help you further.

I am unable to offer this for ab52615 since it has been tested un-suitable for ICC/IF , IHC-P or IP by our lab.

The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

Abcam Scientific Support

Answered on Nov 05 2012

Question

Dear sir/madam,
I am planning to purchase 1852-1 antibodies against androgen receptor through Abcam Europe. I would like to use this antibody for immunohistochemistry on floating brain sections (murine), as one my colleague has already done. However, since on your website you claim that antibody 3165-1 is suitable for immunohistochemistry while 1852-1 is not (although my colleague has used it and it worked) I would like to ask you if it would be possible to get, together with purchased 1852-1 antibody, also a small sample of 3165-1 antibody so I can compare the two which one might be better for our particular application?
Thank you very much in advance!
Yours sincerely,

Abcam community

Verified customer

Asked on Nov 01 2012

Answer

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.

I can strongly recommend to use ab133273 (3165-1) for your IHC-P experiments. This antibody has been tested to work and we therefore can guarantee this for you.

https://www.abcam.com/index.html?datasheet=133273 (or use the following: https://www.abcam.com/index.html?datasheet=133273).

https://www.abcam.com/abpromise

I am sorry to confirm that because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.

I hope this information is helpful. Please do not hesitate to contact us again with any further questions.

Abcam Scientific Support

Answered on Nov 01 2012

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

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

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Datasheets and documents

References

This product has been referenced in:

Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Androgen Receptor antibody [EPR1535(2)]