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17-10489

Sigma-Aldrich

ChIPAb+ Androgen Receptor- ChIP Validated Antibody and Primer Set

from rabbit, purified by affinity chromatography

Synonym(s):

Dihydrotestosterone receptor, Nuclear receptor subfamily 3 group C member 4, DHTR, NR3C4, AR

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.32

biological source

rabbit

Quality Level

clone

polyclonal

purified by

affinity chromatography

species reactivity

rat, amphibian, bovine, human, mouse, chicken

manufacturer/tradename

ChIPAb+
Upstate®

technique(s)

ChIP: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

NCBI accession no.

UniProt accession no.

shipped in

dry ice

General description

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.
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.

Specificity

Recognizes Androgen Receptor. No cross-reactivity with estrogen, progesterone or glucocorticoid receptors.

Immunogen

Peptide corresponding to amino acids 1-21 of the Human Androgen Receptor (MEVQLGLGRVYPRPPSKTYRG)

Application

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.
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.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Transcription Factors

Packaging

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

Quality

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.

Physical form

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’
ImmunoAffinity Purified

Storage and Stability

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.

Analysis Note

Control
Includes normal rabbit IgG and primers specific for prostate-specific antigen (PSA) enhancer region.

Legal Information

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

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 Class Code

10 - Combustible liquids


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Xuanrong Chen et al.
Cell death & disease, 13(11), 927-927 (2022-11-06)
Androgen receptor (AR) plays an important role in the progression of prostate cancer and has been targeted by castration or AR-antagonists. The emergence of castration-resistant prostate cancer (CRPC) after androgen deprivation therapy (ADT) is inevitable. However, it is not entirely
Pasquale Nigro et al.
Diabetes, 70(6), 1250-1264 (2021-02-11)
Recent studies demonstrate that adaptations to white adipose tissue (WAT) are important components of the beneficial effects of exercise training on metabolic health. Exercise training favorably alters the phenotype of subcutaneous inguinal WAT (iWAT) in male mice, including decreasing fat
Xiangnan Guan et al.
Nature, 606(7915), 791-796 (2022-03-25)
Immune checkpoint blockade has revolutionized the field of oncology, inducing durable anti-tumour immunity in solid tumours. In patients with advanced prostate cancer, immunotherapy treatments have largely failed1-5. Androgen deprivation therapy is classically administered in these patients to inhibit tumour cell
Arnaud Blomme et al.
Nature communications, 11(1), 2508-2508 (2020-05-20)
Despite the clinical success of Androgen Receptor (AR)-targeted therapies, reactivation of AR signalling remains the main driver of castration-resistant prostate cancer (CRPC) progression. In this study, we perform a comprehensive unbiased characterisation of LNCaP cells chronically exposed to multiple AR
Alessandro Romanel et al.
Nature communications, 8(1), 48-48 (2017-07-01)
Prostate cancer is a highly heritable molecularly and clinically heterogeneous disease. To discover germline events involved in prostate cancer predisposition, we develop a computational approach to nominate heritable facilitators of somatic genomic events in the context of the androgen receptor

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