中文摘要：

目的以原核表达的小鼠肾上腺蛋白AD-004为免疫原，制备抗AD-004的兔多克隆抗体，并明确其在肾上腺、睾丸组织中的分布。方法将小鼠AD-004cDNA克隆于原核表达载体PET28，经IPTG诱导AD-004的表达。用Ni^2＋-NTA纯化后的蛋白为免疫原制备抗AD-004多克隆抗体。通过Western印迹鉴定抗体特异性及效价，并用此抗体对小鼠睾丸、肾上腺组织进行免疫组织化学分析。结果此原核表达系统可高效表达AD-004蛋白，Western印迹结果显示所制备的抗体具有很高的效价和特异性，免疫组化提示AD-004蛋白在肾上腺的髓质表达较高，而在睾丸中主要表达于间质的Leydig细胞。结论本研究获得了小鼠AD-004原核表达蛋白，成功制备了AD-004多克隆抗体，并发现其在睾丸间质Leydig细胞表达最高，提示AD-004可能在睾丸性激素的合成过程中起重要作用。

英文摘要：

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 IPTG and then purified by Ni^2＋ -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.