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17-10489 Sigma-AldrichChIPAb+ Androgen Receptor- ChIP Validated Antibody and Primer Set

ChIPAb+ &rogen Receptor set includes the &rogen Receptor antibody, a Normal Rabbit IgG, & control primers, which amplify a 85 bp region of the human prostate specific antigen (PSA) enhancer region.

ChIPAb+ Androgen Receptor- ChIP Validated Antibody and Primer Set MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

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Key Applications: Chromatin Immunoprecipitation (ChIP), Western Blotting, Immunohistochemistry, Immunoprecipitation Research Category: Epigenetics & Nuclear Function Research Sub-Category: Transcription Factors

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Key Spec Table

Species Reactivity	Key Applications
H, M, R, Am, B, Ch	ChIP, WB, IHC, IP

Description
Catalogue Number	17-10489
Brand Family	Upstate
Trade Name	ChIPAb+ Upstate
Description	ChIPAb+ Androgen Receptor- ChIP Validated Antibody and Primer Set
Overview	All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction. The ChIPAb+™ Androgen Receptor set includes the Androgen Receptor antibody, a Normal Rabbit IgG, and control primers which amplify a 85 bp region of the human prostate specific antigen (PSA) enhancer region. The Androgen Receptor antibody and negative controls are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of Androgen Receptor associated chromatin.
Alternate Names	Dihydrotestosterone receptor Nuclear receptor subfamily 3 group C member 4 DHTR NR3C4 AR
Background Information	The androgen receptor (AR) is a type of nuclear receptor which is activated by binding of either of the androgenic hormones testosterone or dihydrotestosterone. The main function of the androgen receptor is as a DNA binding transcription factor which regulates gene expression. However the androgen receptor also has additional functions independent of DNA binding. The androgen receptor is most closely related to the progesterone receptor, and progestins in higher dosages can block the androgen receptor.

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Product Information
Format	Affinity Purified
Control	Includes normal rabbit IgG and primers specific for prostate-specific antigen (PSA) enhancer region.
Presentation	Anti-Androgen Receptor (Rabbit Polyclonal), Part No. CS207339. One vial containing 125 µL (0.2 µg/ µL) purified rabbit polyclonal in buffer containing 0.1 M Tris-Glycine (pH7.4),150mM NaCl, and 0.05% sodium azide, before the addition of 30% glycerol. Store at -20°C. Normal Rabbit IgG, Part No. PP64B. One vial containing 125 µg (1.0 µg/ µL) Rabbit IgG in 125 µL storage buffer containing 0.05% sodium azide. Store at -20°C. ChIP Primers, PSA. Part No. CS207340. One vial containing 75 μL of 5 μM of each primer specific for human prostate-specific antigen enhancer region. (chr19:51354179+51354263, hg19 build) Store at -20°C. FOR: 5’ GCC TGG ATC TGA GAG AGA TAT CAT C 3’ REV: 5’ ACA CCT TTT TTT TTC TGG ATT GTT G 3’
Quality Level	MQ100

Applications
Application	ChIPAb+ &rogen Receptor set includes the &rogen Receptor antibody, a Normal Rabbit IgG, & control primers, which amplify a 85 bp region of the human prostate specific antigen (PSA) enhancer region.
Key Applications	Chromatin Immunoprecipitation (ChIP) Western Blotting Immunohistochemistry Immunoprecipitation
Application Notes	Chromatin Immunoprecipitation: LNCaP cells were hormone starved for 72 hours, then induced overnight with 1 µM CI-4AS-1. ChIP was performed using ~1e5 cell equivalents per IP using 1 µg of either Normal Rabbit IgG (Part No. PP64B), or 1 µg Anti-Androgen Receptor (Part No.CS207339) and the Magna ChIP™ HiSens (Cat. # 17-10460). Successful immunoprecipitation of androgen receptor associated DNA fragments was verified by qPCR using ChIP Primers, PSA (Part No. CS207340) as a positive locus, and GAPDH Promoter (Part No. 22-004) as a negative locus. Data are presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated. Please refer to the Magna ChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens(Cat. # 17-10461) protocol for experimental details. Chromatin Immunoprecipitation: LNCaP cells were hormone starved for 72 hours, then induced overnight with vehicle (untreated) or 1 µM CI-4AS-1 (treated). ChIP was performed using ~ 1e5 cell equivalents per IP using 3 µg of either Normal Rabbit IgG (Part No.PP64B ), or 3 µg Anti-Androgen Receptor (Part No.CS207339) and the Magna ChIP™ HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of androgen receptor associated DNA fragments was verified by qPCR using ChIP Primers, PSA (Part No. CS207340) and the negative locus was the HNRP2AB1 primer set. Delta delta Ct calculations reporting fold enrichment at the positive locus over the negative locus are reported. Please refer to the MagnaChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens (Cat. # 17-10461) protocol for experimental details. Western Blot Analysis: A 1:200-1:500 dilution of this antibody detected Androgen Receptor in RIPA lysates from 20 μg of human LNCaP cells. Immunoprecipitation: A representative lot immunoprecipitated Androgen Receptor from 500 μg of LNCaP RIPA lysate. Immunocytochemistry: A 1:50 dilution of a representative lot showed positive immunostaining for Androgen Receptor in LNCaP cells fixed with 2% formaldehyde/5% acetic acid and permeabilized with 0.1% Triton X-100.

Biological Information
Immunogen	Peptide corresponding to amino acids 1-21 of the Human Androgen Receptor (MEVQLGLGRVYPRPPSKTYRG)
Host	Rabbit
Specificity	Recognizes Androgen Receptor. No cross-reactivity with estrogen, progesterone or glucocorticoid receptors.
Species Reactivity	Human Mouse Rat Amphibian Bovine Chicken
Antibody Type	Polyclonal Antibody
Entrez Gene Number	NP_000035
Entrez Gene Summary	The androgen receptor gene is more than 90 kb long and codes for a protein that has 3 major functional domains: the N-terminal domain, DNA-binding domain, and androgen-binding domain. The protein functions as a steroid-hormone activated transcription factor. Upon binding the hormone ligand, the receptor dissociates from accessory proteins, translocates into the nucleus, dimerizes, and then stimulates transcription of androgen responsive genes. This gene contains 2 polymorphic trinucleotide repeat segments that encode polyglutamine and polyglycine tracts in the N-terminal transactivation domain of its protein. Expansion of the polyglutamine tract causes spinal bulbar muscular atrophy (Kennedy disease). Mutations in this gene are also associated with complete androgen insensitivity (CAIS). Two alternatively spliced variants encoding distinct isoforms have been described.
Gene Symbol	AR DHTR SMAX1 HUMARA KD SBMA TFM OTTHUMP00000061928 AIS NR3C4
Purification Method	ImmunoAffinity Purified
UniProt Number	P10275

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

Product Usage Statements
Quality Assurance	Chromatin Immunoprecipitation: Sonicated chromatin prepared from hormone starved (72H), then induced (1 µM CI-4AS-1) overnight LNCaP cells (~1e5 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of either Normal Rabbit IgG (Part No. PP64B), or 1 µg Anti-Androgen Receptor (Part No.CS207339) and the Magna ChIP™ HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of androgen receptor associated DNA fragments was verified by qPCR using ChIP Primers, PSA (Part No. CS207340). Please refer to the MagnaChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens (Cat. # 17-10461) protocol for experimental details.
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Product Usage Statements

Quality Assurance

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from hormone starved (72H), then induced (1 µM CI-4AS-1) overnight LNCaP cells (~1e5 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 1 µg of either Normal Rabbit IgG (Part No. PP64B), or 1 µg Anti-Androgen Receptor (Part No.CS207339) and the Magna ChIP™ HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of androgen receptor associated DNA fragments was verified by qPCR using ChIP Primers, PSA (Part No. CS207340).
Please refer to the MagnaChIP™ HiSens (Cat. # 17-10460) or EZ-MagnaChIP™ HiSens (Cat. # 17-10461) protocol for experimental details.

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Packaging Information
Material Size	25 assays
Material Package	25 assays per set. Recommended use: 1 µg of antibody per chromatin immunoprecipitation (dependent upon biological context).

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Catalogue Number	GTIN
17-10489	04053252922466

Documentation

ChIPAb+ Androgen Receptor- ChIP Validated Antibody and Primer Set MSDS

Title
Safety Data Sheet (SDS)

ChIPAb+ Androgen Receptor- ChIP Validated Antibody and Primer Set Certificates of Analysis

Title	Lot Number
ChIPAb+ Androgen Receptor	2465311
ChIPAb+ Androgen Receptor - 3083466A	3083466A
ChIPAb+ Androgen Receptor - 3201926	3201926
ChIPAb+ Androgen Receptor - 3245808	3245808
ChIPAb+ Androgen Receptor - 3474388	3474388
ChIPAb+ Androgen Receptor - 3689486	3689486
ChIPAb+ Androgen Receptor - 3762793	3762793
ChIPAb+ Androgen Receptor - 3856703	3856703
ChIPAb+ Androgen Receptor - 3982021	3982021
ChIPAb+ Androgen Receptor - 4118949	4118949

References

Reference overview	Pub Med ID
Genome-wide analysis of androgen receptor binding sites in prostate cancer cells. Cheng, Y; Yu, P; Duan, X; Liu, C; Xu, S; Chen, Y; Tan, Y; Qiang, Y; Shen, J; Tao, Z Exp Ther Med 9 2319-2324 2015 Show Abstract The transformation of prostate cancer from an androgen-dependent state to an androgen-independent state is a lethal progression. Alterations in transcriptional programs are the basis of prostate cancer deterioration. The androgen receptor (AR), a member of the nuclear hormone receptor superfamily, mediates prostate cancer progression by functioning primarily through the ligand-activated transcription of target genes. Therefore, a detailed map of AR-regulated genes and AR genomic binding sites is required for hormone-naive and castration-resistant prostate cancer. Through the use of chromatin immunoprecipitation in combination with direct sequencing, 4,143 AR binding sites were defined in the LNCaP androgen-sensitive prostate cancer cell line. Using the same method, 2,380 AR binding regions were identified in the LNCaP-AI long-term androgen-deprived cell line. Approximately 8.5% (354/4,143) of the binding regions were mapped to within 2 kb of the transcription start site (TSS) in the LNCaP cells, while ∼12.6% (299/2,380) were mapped to within 2 kb of the TSS in the LNCaP-AI cells. In total, the study mapped 2,796 genes in LNCaP cells and 1,854 genes in LNCaP-AI cells. The cell lines shared 789 mutual genes. In addition, gene ontology (GO) analysis of the genes revealed that there was a notable overlap between the GO terms in the LNCaP cells and LNCaP-AI cells. However, GO terms within the biological process domain that were only observed in the LNCaP-AI cells included the reproduction process, death, immune system process, multi-organism process, pigmentation and viral reproduction. The major genes in the different GO terms were TNFAIP8, RTN4, APP and SYNE1. Through analyzing the AR binding sites in the two cell types, the present study aimed to map potential AR-regulated genes, identify their associated transcription factors and provide a new perspective on the biological processes in the development of prostate cancer. The results provided a valuable data set that furthered the understanding of the genome-wide analysis of AR binding sites in prostate cancer cells, which may be exploited for the development of novel prostate cancer therapeutic strategies.	26136980

Reference overview

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