van Laar J H, Voorhorst-Ogink M M, Zegers N D, Boersma W J, Claassen E, van der Korput J A, Ruizeveld de Winter J A, van der Kwast T H, Mulder E, Trapman J
Department of Biochemistry II, Erasmus University Rotterdam, The Netherlands.
Mol Cell Endocrinol. 1989 Nov;67(1):29-38. doi: 10.1016/0303-7207(89)90227-x.
Antibodies against the N-terminal domain of the human androgen receptor (hAR) were prepared by two different approaches. Firstly, rabbits were immunized with a beta-galactosidase-hAR (amino acids (aa) 174-353) fusion protein. Secondly, two synthetic peptides corresponding to potentially antigenic sites located within this fragment (aa 201-222 and 301-320) were used as immunogens. The obtained antisera contained high titer anti-hAR antibodies as was established with several independent methods (e.g. sucrose gradient centrifugation, immunoprecipitation, Western blotting). The two anti-peptide antisera specifically stained nuclei of glandular epithelial cells in frozen sections of human prostate tissue. Progesterone, estradiol and glucocorticoid receptors were not immunoprecipitated with these antisera. The specific hAR antibodies provide new tools for the characterization of this steroid receptor as well as for diagnostic purposes in pathology of the human prostate and androgen resistance.
通过两种不同方法制备了针对人雄激素受体（hAR）N端结构域的抗体。首先，用β-半乳糖苷酶-hAR（氨基酸（aa）174 - 353）融合蛋白免疫兔子。其次，将对应于该片段内潜在抗原位点的两种合成肽（aa 201 - 222和301 - 320）用作免疫原。通过几种独立方法（如蔗糖梯度离心、免疫沉淀、蛋白质免疫印迹法）确定，所获得的抗血清含有高滴度的抗hAR抗体。两种抗肽抗血清在人前列腺组织冰冻切片中特异性地染色腺上皮细胞核。孕酮、雌二醇和糖皮质激素受体不能被这些抗血清免疫沉淀。特异性hAR抗体为该类固醇受体的特性研究以及人类前列腺病理学和雄激素抵抗的诊断目的提供了新工具。
