14-3-3ζ蛋白单克隆抗体的制备、鉴定及初步应用  

Preparation and preliminary application of anti-14-3-3ζ monoclonal antibody

作　　者：董长松[1,2] 李尧华[1] 叶懿文[1] 李昕[1] 于顺[1] 杨慧[3] 陈彪[1] 

机构地区：[1]首都医科大学宣武医院老年病研究所神经生物学研究室,教育部神经变性病重点实验室,北京100053 [2]吉林医药学院附属医院消化科,吉林132013 [3]首都医科大学神经科学研究所,北京100053

出　　处：《神经解剖学杂志》2009年第6期631-634,共4页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金重点(30430280,30271437,30270482);北京市自然科学基金(7022011)资助项目

摘　　要：为了探讨14-3-3ζ蛋白在中枢神经系统内的分布,本文采用纯化的基因重组型14-3-3ζ蛋白免疫Balb/c小鼠,用杂交瘤技术制备抗14-3-3ζ蛋白单克隆抗体;Western blot及免疫组织化学法鉴定抗体。结果显示:制备的单克隆抗体能特异性识别基因重组14-3-3ζ蛋白和大鼠脑匀浆32kD蛋白。免疫组化染色结果显示,14-3-3ζ蛋白在大鼠中枢神经系统广泛分布,在皮层、海马、杏仁核和基底前脑的免疫阳性染色较强,免疫阳性产物位于神经元胞浆内。本研究结果提示,14-3-3ζ蛋白可能具有多样化的生理功能。In order to identify the distribution of 14-3-3ζ protein in the central nervous system, 14-3-3ζ monoclonal antibody was prepared. Purified recombinant 14-3-3ζ protein was used to immunize Balb/e mouse, and hybfidoma technique was used to screen a cells line, which released the target antibody. Western blot and immunohistochemical technique were used to identify the antibody. The resluts showed that the prepared antibody could recognise recombinant 14-3-3ζ protein and the approximately 32 kD protein in the rat brain homogenize. Immunohistochemical staining results showed that the 14-3-3ζ protein was widely distributed in the rat central nervous system, especially in the cortex, hippocampus, amygdala and basal forebrain. 14-3-3ζ immunoreactivity was mainly in the cytoplasm of neurons. The present results indicate that 14-3-3ζ protein could be have various physiological function.

关 键 词：14-3-3ζ蛋白 单克隆抗体 中枢神经系统 大鼠 

分 类 号：R392.1[医药卫生—免疫学]