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草鱼细胞内凋亡抑制蛋白多克隆抗体的制备和检测

发表时间:2013-09-15  浏览量:941  下载量:396
全部作者: 张小宁,邱倩,李慧,黄德军,张迎梅
作者单位: 兰州大学生命科学学院
摘 要: 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 blotting可以检测到IAP在草鱼肝脏组织中的表达。结论:研究得到的IAP多克隆抗体可用于草鱼IAP表达检测、亚细胞定位及抗凋亡功能研究等。
关 键 词: 动物学;多克隆抗体;原核表达;凋亡抑制蛋白;草鱼
Title: Preparation and assay of polyclonal antibody for inhibitor of apoptosis protein in Ctenopharyngodonidellus
Author: ZHANG Xiaoning, QIU Qian, LI Hui, HUANG Dejun, ZHANG Yingmei
Organization: School of Life Sciences, Lanzhou University
Abstract: Objective: The present study aims to make the polyclonal antibody for inhibitor of apoptosis protein (IAP) in Ctenopharyngodonidellus. Methods: We designed a pair of specific primers for IAP gene in C.idellus using bioinformatics technology, amplified a 996 bp 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.
Key words: zoology; polyclonal antibody; prokaryotic expression; inhibitor of apoptosis protein; Ctenopharyngodonidellus
发表期数: 2013年9月第17期
引用格式: 张小宁,邱倩,李慧,等. 草鱼细胞内凋亡抑制蛋白多克隆抗体的制备和检测[J]. 中国科技论文在线精品论文,2013,6(17):1651-1656.
 
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