Revision 5
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, W-S, IP, ChIP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

80

Source/Isotype:

Rabbit IgG

UniProt ID:

#P10275-3

Entrez-Gene Id:

367

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Immunoprecipitation 1:200
Chromatin IP 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Androgen Receptor (AR-V7 Specific) (E3O8L) Rabbit mAb recognizes endogenous levels of total AR-V7 protein. This antibody does not cross-react with full-length AR protein, but immunoprecipitates a 120 kDa protein of unknown identity.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu639 of human androgen receptor (V7 isoform) protein.

Background

Androgen receptor (AR), a zinc finger transcription factor belonging to the nuclear receptor superfamily, is activated by phosphorylation and dimerization upon ligand binding (1). This promotes nuclear localization and binding of AR to androgen response elements in androgen target genes. Research studies have shown that AR plays a crucial role in several stages of male development and the progression of prostate cancer (2,3).
The AR3 or AR-V7 isoform, which lacks the typical ligand binding domain, is created through the alternative splicing of cryptic exons (4-5). AR-V7 is frequently expressed in castration-resistant prostate cancer (CRPC) and while dependent on the activity of the full-length androgen receptor (AR-FL), AR-V7 can activate a completely distinct transcriptional program (6-8). While enzalutamide and abiraterone have been beneficial in treating CRPC through the ligand binding domain of AR-FL, resistance in patients has been shown to be associated with AR-V7 detection in circulating tumor cells (9-12). Studies probing into mechanisms of overcoming this resistance are currently being explored and may help in stratifying patient populations for more personalized therapies (13-15).

  1. Li, J. and Al-Azzawi, F. (2009) Maturitas 63, 142-8.
  2. Avila, D.M. et al. (2001) J. Steroid. Biochem. Mol. Biol. 76, 135-142.
  3. Montgomery, J.S. et al. (2001) J. Pathol. 195, 138-146.
  4. Hu, R. et al. (2009) Cancer Res 69, 16-22.
  5. Guo, Z. et al. (2009) Cancer Res 69, 2305-13.
  6. Watson, P.A. et al. (2010) Proc Natl Acad Sci U S A 107, 16759-65.
  7. Sun, S. et al. (2010) J Clin Invest 120, 2715-30.
  8. Hu, R. et al. (2012) Cancer Res 72, 3457-62.
  9. Scher, H.I. et al. (2012) N Engl J Med 367, 1187-97.
  10. de Bono, J.S. et al. (2011) N Engl J Med 364, 1995-2005.
  11. Ryan, C.J. et al. (2013) N Engl J Med 368, 138-48.
  12. Antonarakis, E.S. et al. (2014) N Engl J Med 371, 1028-38.
  13. Liu, C. et al. (2014) Clin Cancer Res 20, 3198-3210.
  14. Sarwar, M. et al. (2016) Oncotarget 7, 63065-63081.
  15. Ku, S.Y. et al. (2017) Science 355, 78-83.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation ChIP: Chromatin IP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
CST is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 5
#19672

Androgen Receptor (AR-V7 Specific) (E3O8L) Rabbit mAb

Western Blotting Image 1: Androgen Receptor (AR-V7 Specific) (E3O8L) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using Androgen Receptor (AR-V7 Specific) (E3O8L) Rabbit mAb (upper) and Β-Actin (D6A8) Rabbit mAb #8457 (lower). Signal corresponding to the androgen receptor V7 isoform is detected in 22Rv1 and VCAP cells (AR-V7-positive), but is not detected in LNCaP and DU 145 cells (AR-V7-negative), confirming specificity of the antibody.
Western Blotting Image 1: Androgen Receptor (AR-V7 Specific) (E3O8L) Rabbit mAb Expand Image
Simple Western™ analysis of lysates (1 mg/mL) from 22Rv-1 cells using Androgen Receptor (AR-V7 Specific) (E3O8L) Rabbit mAb #19672. The virtual lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Immunoprecipitation Image 1: Androgen Receptor (AR-V7 Specific) (E3O8L) Rabbit mAb Expand Image
Immunoprecipitation of AR-V7 from 22Rv1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb XP® Isotype Control #3900, and lane 3 is Androgen Receptor (AR-V7 Specific) (E3O8L) Rabbit mAb. Western blot analysis was performed using Androgen Receptor (AR-V7) (E3O8L) Rabbit mAb.
Chromatin Immunoprecipitation Image 1: Androgen Receptor (AR-V7 Specific) (E3O8L) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from 22-RV-1 cells, treated with 10 nM DHT for 4hr and either Androgen Receptor (AR-V7 Specific) (E3O8L) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SPINK5 upstream primers, ABCB5 upstream primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.