合成多肽制备的抗体用于蛋白质免疫组织细胞化学定位研究  

Protein Immunocytochemistry Localization by Using Antibody Prepared With Synthesized Polypeptide

作　　者：邢立静[1] 种康[1] 许智宏[1] 王锐[2] 谭克辉[1] 

机构地区：[1]中国科学院植物研究所,中国科学院光合作用与环境分子生理学重点实验室,北京100093 [2]兰州大学生命科学学院,兰州730000

出　　处：《生物化学与生物物理进展》2004年第2期150-153,共4页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金资助项目 ( 3 0 170 0 94);中国科学院知识创新重大项目资助 (KSCX2 SW 3 0 5 )~~

摘　　要：对本实验室克隆的小麦春化相关基因VER2编码蛋白的氨基酸序列进行软件分析 ,确定了具有免疫原性的一段 (2 0 4～ 2 1 5位 )含 1 2个氨基酸的短肽 ,用化学法合成了该短肽并将其与牛血清白蛋白 (BSA)相偶联 ,用偶联物作为抗原免疫家兔获得该 1 2肽的抗体 .分别用VER2全蛋白抗体和该 1 2肽抗体对VER2蛋白进行免疫细胞定位比较分析 ,两者所得结果一致 ,即二者在免疫组化检测蛋白质时的灵敏性和特异性方面具有一致的特性 .结果表明 ,化学法合成短肽制备的抗体可用于蛋白质的免疫组织细胞化学分析 ,该方法快捷灵敏 。The deduced amino acid sequence of the VER2 gene was analyzed in a computer. A peptide corresponding to 204～215 amino acids in VER2 sequence was synthesized by chemical method and purified using HPLC. The polypeptide was conjugated with BSA and the covalent complex was used as antigen to immunize rabbits. The antibody was then affinity purified before being used to probe VER2 protein in Paraplast sections. The immunolocalization results detected by the peptide antibody were consistent with that of the full length VER2 protein antibody. It is concluded that antibody prepared with synthetic peptide is credible and sensitive enough for immune labeling of proteins in paraplast sections. It is a sensitive and effective approach to combine preparation of antibody from synthetic peptide with paraplast section production for protein immunocytochemistry study.

关 键 词：化学合成短肽 VER2蛋白 抗体纯化 免疫组织细胞化学 植物体 

分 类 号：Q946.1[生物学—植物学] Q945