androgen receptor Recombinant antibody

androgen receptor Recombinant Antibody for IF, IHC, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

Human

Applications

WB, IHC, IF, ELISA

Conjugate

Unconjugated

CloneNo.

4O18

Cat no : 81844-1-RR

Synonyms

AIS, androgen receptor, AR, DHTR, Dihydrotestosterone receptor, HUMARA, HYSP1, KD, NR3C4, SBMA, SMAX1, TFM

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Validation Data Gallery

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  WB analysis using 81844-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 81844-1-RR (androgen receptor antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis using 81844-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 81844-1-RR (androgen receptor antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.

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  IHC staining of human prostate cancer using 81844-1-RR

  Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 81844-1-RR (androgen receptor antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human prostate cancer using 81844-1-RR

  Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue slide using 81844-1-RR (androgen receptor antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of LNCaP using 81844-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed LNCaP cells using androgen receptor antibody (81844-1-RR, Clone: 4O18 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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Tested Applications

• Positive WB detected in: LNCaP cells, MCF-7 cells, K-562 cells
• Positive IHC detected in: human prostate cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF detected in: LNCaP cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF): IF : 1:500-1:2000

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

81844-1-RR targets androgen receptor in WB, IHC, IF, ELISA applications and shows reactivity with Human samples.

• Tested Reactivity: Human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: androgen receptor fusion protein Ag17291
• Full Name: androgen receptor
• Calculated Molecular Weight: 914 aa, 99 kDa
• Observed Molecular Weight: 110-120 kD
• GenBank Accession Number: BC132975
• Gene Symbol: AR
• Gene ID (NCBI): 367
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Androgen receptor (AR) is a steroid hormone receptor for androgenic hormones such as 17β-Hydroxy-3-oxo-4-androstene and DHT. AR plays a vital role in developing and maintaining male sex phenotypes as well as an additional role in regulating bone metabolism.

1. What is the molecular weight of AR? Are there any isoforms of AR?
The molecular weight of full-length androgen receptor (AR-B) is 110 kDa. An additional variant, AR-A, has an 87 kDa size and lacks the N-terminal 187 amino acids of AR-A (PMID: 8108393). Recently, more splice variants of AR have been discovered, raising protein products of around 80 kDa length (PMID: 19244107), as well as an AR45 variant of 45 kDa size (PMID: 15634333). AR splice variants differ in their cell line-specific expression (PMID: 24570075).

2. What is the subcellular localization of AR?
AR can be present in either or both of the cytoplasm and nucleus. In androgen-deprived cells, AR is found predominantly in the cytoplasm, while stimulation by androgens causes enrichment of androgen-bound AR in the nucleus. AR shuttles between the cytoplasm and nucleus and its phosphorylation state has an impact on subcellular localization (PMID: 16282370).

3. Is AR post-translationally modified?
Post-translational modifications of the AR include phosphorylation, acetylation, methylation, SUMOylation, and ubiquitination (PMID: 21820033). These modifications have an impact on receptor stability, activity, and can change the observed molecular weight of the AR.

4. How to study AR signaling in cell culture?
It is important to control levels of cell stimulation while also looking at AR signaling. Fetal bovine serum (FBS) that is typically used in cell culture contains low levels of 17β-Hydroxy-3-oxo-4-androstene that are enough to stimulate the growth of prostate cells (PMID: 19676093), including the LNCaP cell line that is a commonly used human prostatic carcinoma cell model (PMID: 6831420). One possibility for complete 17β-Hydroxy-3-oxo-4-androstene deprivation is to use charcoal stripped FBS that removes lipophilic agents, including androgens. It is also not recommended to use phenol red in your medium because it is a weak estrogen (PMID: 3458212). Cell stimulation is often conducted by DHT.

5. What is the role of AR in prostate cancer?
AR plays a key role in the development and physiology of the prostate gland, and also cancer progression (PMID: 15082523). Mutations in AR altering ligands have been observed. The progression of the prostate cancer depends on AR activity and therefore blocking AR activity or lowering androgen levels is a key step related to androgen deprivation therapy (ADT).