Protein expression and preparation of polyclonal antibody of AD-004 and study on its expression in the adrenal and testis
- VernacularTitle:AD-004蛋白表达、抗体制备及其在睾丸和肾上腺组织中的表达分析
- Author:
Jie QIAO
;
Cuixia ZHENG
;
Xia CHEN
;
Xuesong LI
;
Shengxian LI
;
Qinyun MA
;
Shuangxia ZHAO
;
Guoyue YUAN
;
Mingdao CHEN
- Publication Type:Journal Article
- Keywords:
Protein AD-004;
Prokaryotic expression;
Adenylate kinases;
Antibodies, polyclonal
- From:
Chinese Journal of Endocrinology and Metabolism
1986;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare rabbit antibody against mouse AD-004 by AD-004 expressed in the prokaryotic expression system and to identify its distribution in the testis and adrenal. Methods The full-length cDNA of mouse AD-004 was cloned into PET28 plasmid, and the protein was induced in E. coli BL21 bacteria by adding IPTC and then purified by Ni2+ -NTA column. The purified protein was used as an immunogen to prepare polyclonal antibody ( pAb) of AD-004. The specificity of the antibody was detected by Western blotting. Immunohistochemical staining was performed in the mouse adrenal and testis via pAb of AD-004. Results Hisfused AD-004 was expressed efficiently in the prokaryotic system. Western blot analysis showed that the polyclonal antibody was duly bound to purified AD-004 with high specificity and sensitivity. AD-004 could be abundantly identified in the adrenal medulla and mainly expressed in the Leydig cells of testis. Conclusion The mouse protein of AD-004 is obtained from the prokaryotic expression system. The rabbit anti-AD-004 antibody has been prepared successfully. AD-004 protein is mainly localized in the interstitium of testis, suggesting that AD-004 may play a role in the synthesis of sex-steroid hormone.
