BACKGROUND
The androgen receptor (AR) mediating androgen effects is a member of the steroid and thyroid hormone receptor gene superfamily encoding ligand-dependent nuclear transcription factors (reviewed in 1). Based on their functions, the AR molecule can be sub-divided into three major domains, an N-terminal domain which modulates transcription efficiency, a central DNA-binding domain which binds to a target gene hormone response element, and a C-terminal hormone-binding domain which binds androgens. Mutations in the AR gene are associated with androgen insensitivity syndrome, a disorder that causes XY genotypic males to develop as phenotypic females because of their inability to respond to androgens (reviewed in 2). AR mutations have also been identified in human prostate cancer specimens and the LNCaP human prostate carcinoma cell line.2 Monoclonal and polyclonal antibodies to the AR have identified AR-positive cells in a variety of tissues including male and female sexual organs, kidney, liver, adrenal cortex, pituitary gland, skeletal, cardiac, and smooth muscle cells.3 In SDS-PAGE, the androgen receptor migrates at a reduced molecular weight of ~100 kD.
PharMingen currently offers four monoclonal antibodies directed against the human androgen receptor for various applications.
G122-25.3 (cat. #15051A) recognizes an epitope localized within the DNA-binding domain (between amino acids 486 and 651) of the human androgen receptor. The antibody does not recognize estrogen or progesterone receptors. Purified recombinant human androgen receptor protein was used as immunogen.4 Applications include western blot analysis (2µg/ml) immunoprecipitation (1-2 µg/one million cells) , flow cytometry (titrate between 0.1 and 4 µg/one million cells), and immunohistochemical staining of frozen and paraffin-embedded tissue sections (titrate between 5 and 20 µg/ml). This mAb also stains rat AR in frozen prostat tissue sections.
G122-434 (cat. #15061A) recognizes an epitope localized within the N-terminal domain (between amino acids 33 and 485) of the human androgen receptor. The antibody does not recognize estrogen or progesterone receptors. Purified recombinant human androgen receptor protein was used as immunogen.4 This antibody is optimal for use in western blot analysis (2µg/ml). Other applications include immunoprecipitation (1-2 µg/one million cells), and immunohistochemical staining of frozen tissue sections (titrate between 10 and 20 µg/ml).
G122-77 (cat. #15071A) recognizes an epitope localized within the DNA-binding domain (between amino acids 486 and 651) of the human androgen receptor. The antibody does not recognize estrogen or progesterone receptors. Purified recombinant human androgen receptor protein was used as immunogen.4 Applications include immunoprecipitation (1-2 µg/one million cells), and immunohistochemical staining of frozen tissue sections (titrate between 10 and 20 µg/ml). This mAb also stains rat AR in frozen prostate tissue sections.
G122-238 (cat. #65461A) recognizes an epitope localized within the N-terminal domain (between amino acids 33 and 485) of the human androgen receptor. The antibody does not recognize estrogen or progesterone receptors. Purified recombinant human androgen receptor protein was used as immunogen.4 Applications include immunohistochemical staining of frozen and paraffin-embedded tissue sections (titrate between 5 and 20 µg/ml). Clone G122-238 can also be used for western blot analysis, however Clone G122-434 (cat. #15061A) is our optimal clone for western blot analysis.
| Cat. No. | Description | Clone | Size |
| 15051A | Purified anti-human Androgen Receptor (a.a. 486-651) | G122-25.3 | 0.1 mg |
| 15061A | Purified anti-human Androgen Receptor (a.a. 33-485) | G122-434 | 0.1 mg |
| 15071A | Purified anti-human Androgen Receptor (a.a. 486-651) | G122-77 | 0.1 mg |
| 65461A | Purified anti-human Androgen Receptor (a.a. 33-485) | G122-238 | 0.1 mg |
REFERENCES
1. Fuller, P.J. 1991. The steroid receptor superfamily: mechanisms of diversity.
FASEB
5:3092-3099.
2. Newmark, J.R., D.O. Hardy, D.C. Tonb, B.S. Carter, J.I. Epstein, W.B. Isaacs, T.R. Brown, and E.R. Barrack. 1992. Androgen receptor gene mutations in human prostate cancer.
Proc. Natl. Acad. Sci.
89:6319-6323.
3. Takeda, H., G. Chodak, S. Mutchnik, T. Nakamoto, and C. Chang. 1990. Immunohistochemical localization of androgen receptors with mono- and polyclonal antibodies to androgen receptor.
J. Endocrinology
126:17-25.
4. Chang, C., C. Wang, H.F. DeLuca, T.K. Ross, and C.C.-Y. Shih. 1992. Characterization of human androgen receptor overexpressed in the baculovirus system.
Proc. Natl. Acad. Sci.
89: 5946-5950.
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