草鱼细胞内凋亡抑制蛋白多克隆抗体的制备和检测
Preparation and assay of polyclonal antibody for inhibitor of apoptosis protein in Ctenopharyngodon idellus
目的：制备草鱼细胞内凋亡抑制蛋白（Inhibitor of Apoptosis Protein, IAP）多克隆抗体，为进一步开展IAP蛋白时空差异表达分析、定位和胁迫条件下的抗凋亡功能研究奠定基础。方法：利用生物信息学技术手段设计IAP基因的特异性引物，扩增长度为996 bp的IAP基因片段，运用原核表达系统表达抗原，免疫新西兰大白兔，获得抗血清后，通过亲和纯化得到IAP蛋白特异性的抗体。结果：获得了高效价的IAP蛋白多克隆抗体，Western Blot可以检测到IAP蛋白在草鱼肝脏组织中的表达。结论：本研究得到的IAP多克隆抗体可用于草鱼IAP蛋白表达检测、亚细胞定位及抗凋亡功能研究等。
Objective]: The present study aims to make the polyclonal antibody for inhibitor of apoptosis protein（IAP）in Ctenopharyngodon idellus. [Methods]: We designed a pair of specific primers for IAP gene in C. idellus by bioinformatics technology, amplified a 996bp DNA fragment and introduced this IAP gene fragment into E.coli to express a fusion protein His-IAP which then was injected into New Zealand white rabbits to prepare antiserum. Finally, specific anti-IAP polyclonal antibody was obtained by affinity purification. [Results]: Immunoblot analysis confirmed that IAP protein could be detected in C. idellus liver by this anti-IAP polyclonal antibody. [Conclusion]: An IAP polyclonal antibody is gathered and can be used for expression detection, subcellular localization and anti-apoptotic function research of IAP protein in C. idellus.
李慧、张小宁、邱倩、黄德军、张迎梅
细胞生物学生物化学分子生物学
动物学凋亡抑制蛋白原核表达多克隆抗体草鱼
ZoologyInhibitor of apoptosis proteinProkaryotic expressionPolyclonal antibodyCtenopharyngodon idellus
李慧,张小宁,邱倩,黄德军,张迎梅.草鱼细胞内凋亡抑制蛋白多克隆抗体的制备和检测[EB/OL].(2013-02-05)[2025-08-23].http://www.paper.edu.cn/releasepaper/content/201302-100.点此复制
评论