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61949
Androgen Receptor Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Androgen Receptor Sampler Kit #61949

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Androgen Receptor Sampler Kit: Image 1

Confocal immunofluorescent analysis of LNCaP cells (left, positive) and DU 145 cells (right, negative) using

Androgen Receptor (E3S4N) Rabbit mAb (Carboxy-terminal Antigen) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).

Androgen Receptor Sampler Kit: Image 2

Chromatin immunoprecipitations were performed with cross-linked chromatin from LNCaP cells grown in phenol red free medium and 5% charcoal stripped FBS for 3 d then treated with dihydrotestosterone (DHT, 10 nM) for 4 hours and Androgen Receptor (D6F11) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across KLK2, a known target gene of Androgen Receptor (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

Androgen Receptor Sampler Kit: Image 3

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.

Androgen Receptor Sampler Kit: Image 4

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Androgen Receptor Sampler Kit: Image 5

Immunoprecipitation of Androgen Receptor from LNCaP cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Androgen Receptor (E3S4N) Rabbit mAb (Carboxy-terminal Antigen). Western blot analysis was performed using Androgen Receptor (E3S4N) Rabbit mAb (Carboxy-terminal Antigen). Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.

Androgen Receptor Sampler Kit: Image 6

Chromatin immunoprecipitations were performed with cross-linked chromatin from LNCaP cells grown in phenol red free medium and 5% charcoal stripped FBS for 3 d then treated with dihydrotestosterone (DHT, 10 nM) for 4 hours and either Androgen Receptor (D6F11) XP® 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 SimpleChIP® Human KLK2 Intron 1 Primers #62086, SimpleChIP® Human KLK3 Promoter Primers #32784, 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.

Androgen Receptor Sampler Kit: Image 7

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.

Androgen Receptor Sampler Kit: Image 8

Western blot analysis of extracts from various cell lines using Androgen Receptor (E3S4N) Rabbit mAb (Carboxy-terminal Antigen) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, DU 145 cells are negative for Androgen Receptor expression.

Androgen Receptor Sampler Kit: Image 9

Flow cytometric analysis of DU-145 cells (blue) and LNCaP cells (green) using Androgen Receptor (D6F11) XP® Rabbit mAb (solid lines) or a concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Androgen Receptor Sampler Kit: Image 10

Confocal immunofluorescent analysis of LNCaP (positive, left) and DU145 (negative, right) cells using Androgen Receptor (D6F11) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

Androgen Receptor Sampler Kit: Image 11

Immunohistochemical analysis of paraffin-embedded LNCaP (AR+, left) and DU145 (AR-, right) using Androgen Receptor (D6F11) XP® Rabbit mAb.

Androgen Receptor Sampler Kit: Image 12

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Androgen Receptor (D6F11) XP® Rabbit mAb.

Androgen Receptor Sampler Kit: Image 13

Western blot analysis of extracts from LNCaP (AR+), MCF7 (AR+), PC-3 (AR-), and DU 145 (AR-) cells using Androgen Receptor (D6F11) XP® Rabbit mAb (upper) and β-Actin Antibody #4967 (lower).

To Purchase # 61949T
Product # Size Price Inventory
61949T
1 Kit  (3 x 20 µl)

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Product Description

The Androgen Receptor Antibody Sampler Kit provides an economical means of detecting full-length Androgen Receptor and AR-V7 isoforms. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Androgen Receptor Antibody Sampler kit detects endogenous levels of its target protein. Androgen Receptor (D6F11) XP® Rabbit mAb detects both full-length AR and the AR-V7 isoform. Androgen Receptor (E3S4N) Rabbit mAb (Carboxy-terminal Antigen) detects only full-length AR. Androgen Receptor (AR-V7 Specific) (E3O8L) Rabbit mAb only detects the AR-V7 isoform.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with recombinant protein corresponding to residues near the amino terminal region of human androgen receptor protein, and with synthetic peptides corresponding to residues surrounding Val662 of human androgen receptor protein and 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, 3198-210.
  14. Sarwar, M. et al. (2016) Oncotarget 7, 63065-81.
  15. Ku, S.Y. et al. (2017) Science 355, 78-83.

Pathways & Proteins

Explore pathways + proteins related to this product.

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