蛋白酶体新亚基PSMA8的多克隆抗体制备及鉴定  

Preparation and Characterization of a Poly-clonal Antibody Against a Novel Proteasome Subunit,PSMA8

作　　者：杜伯雨[1] 郭阳[1] 宋伟[2] 

机构地区：[1]湖北医药学院基础医学院,湖北十堰442000 [2]北京协和医学院基础学院中国医学科学院基础医学研究所生化学系,北京100005

出　　处：《湖北医药学院学报》2015年第4期327-332,共6页Journal of Hubei University of Medicine

基　　金：湖北省教育厅自然科学项目(B2014048);湖北医药学院"2011计划"2014年度开放基金创新团队项目(2011JH-2014CXTT06)

摘　　要：目的:制备一种特异的抗PSMA8的多克隆抗体,并可用于PSMA8的鉴定。方法:通过序列比对,找出特异的抗原序列,经过原核表达制备抗原,免疫大鼠后利用Ni+-NTA柱和分子筛层析制备高纯度、高特异的PSMA8多克隆抗体。结果:所制备的抗体可以同PSMA8蛋白特异性结合,此外,PSMA8特异的表达于睾丸组织和精子细胞,而在其他组织和细胞中,包括精原细胞和精母细胞中不表达。结论:本研究制备了高纯度高特异的PSMA8多克隆抗体,可以有效的支持在睾丸组织或精子细胞中对PSMA8功能的研究,并已经成功的应用于动物组织中PSMA8蛋白的免疫组织化学检测。Objective To prepare a specific poly-clonal antibody against PSMA8,which could also be used for PSMA8 charaterizaiton. Methods By using the sequece alignment,target sequence for antigen preparation had been selected. Then the antigen had been prepared and was used to immunize the rats. The poly-clonal antibody had thus been prepared by affinity chromatography and subsequent size-exclusion chromatography. Results The poly-clonal antibody obtained could specifically bind to PSMA8 protein. The results from western blotting experiment had shown that PSMA8 was specifically expressed in testis（ mainly in the sperm）,but not in other tissues and cell lines,including spermatogonia and spermatocytes. Conclusion We had prepared a highly specific poly-clonal antibody against PSMA8. It could be used in the study for further investigation of PSMA8 function within testis tissues and sperm cells.

关 键 词：PSMA8 蛋白酶体 睾丸 

分 类 号：R392[医药卫生—免疫学]