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25015
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet
Primary Antibodies
Antibody Duet

PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet #25015

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PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: Image 1
Western blot analysis of extracts from LNCaP (AR+), VCaP (AR & AR-V7+), 22Rv1 (AR & AR-V7+), and PC-3 (AR-) cells using Androgen Receptor (D6F11) XP® Rabbit mAb (upper), and β-Actin (13E5) Rabbit mAb (lower).
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: Image 2
Western blot analysis of extracts from LNCaP cells grown in phenol red free media and 5% charcoal stripped FBS for 3 d then either untreated (-), dihydrotestosterone (DHT)-treated (+), or DHT-treated and subsequently treated with λ phosphatase and calf intestinal phosphatase (CIP) (+), using Phospho-Androgen Receptor (Ser258) (E4L1I) Rabbit mAb (upper) or Androgen Receptor (D6F11) XP® Rabbit mAb #5153 (lower).
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: Image 3
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).
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: Image 4
Immunoprecipitation of phospho-androgen receptor (Ser258) protein from LNCaP cell extracts. Cells were grown in phenol red free media and 5% charcoal stripped FBS for 3 d then treated with dihydrotestosterone (DHT) (5 nM, 6 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Androgen Receptor (Ser258) (E4L1I) Rabbit mAb. Western blot analysis was performed using Phospho-Androgen Receptor (Ser258) (E4L1I) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: Image 5
Immunohistochemical analysis of paraffin-embedded LNCaP (AR+, left) and DU145 (AR-, right) using Androgen Receptor (D6F11) XP® Rabbit mAb.
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: 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) (5 nM, 6 hr) and either Phospho-Androgen Receptor (Ser258) (E4L1I) 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 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.
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: Image 7
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Androgen Receptor (D6F11) XP® Rabbit mAb.
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: Image 8
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).
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: 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.
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: Image 10
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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #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.
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: Image 11
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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 19 (upper), including KLK2 (lower), a known target gene of Androgen Receptor (see additional figure containing ChIP-qPCR data).
PhosphoPlus® Androgen Receptor (Ser258) Antibody Duet: Image 12
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.
To Purchase # 25015
Cat. # Size Qty. Price
25015S
1 Kit
$ 632

Product Includes Quantity Reactivity MW(kDa) Isotype
Androgen Receptor (D6F11) XP® Rabbit mAb 5153 100 µl H 110 Rabbit IgG
Phospho-Androgen Receptor (Ser258) (E4L1I) Rabbit mAb 16969 100 µl H 110 Rabbit IgG

Product Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.
MW (kDa) 110

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).
Upon ligand binding, AR becomes phosphorylated on many residues, including Ser258 (4).
  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. Gioeli, D. et al. (2002) J Biol Chem 277, 29304-14.

Pathways & Proteins

Explore pathways + proteins related to this product.

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.

For Research Use Only. Not For Use In Diagnostic Procedures.
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