人GrnE单克隆抗体的制备和鉴定  

Preparation and identification of monoclonal antibody against human Grn E

作　　者：李卫玲[1] 张娅玲[1] 李彦青[1] 张丽娟[1] 夏海滨[1] 

机构地区：[1]陕西师范大学生命科学学院,秦巴山区可持续发展协同创新中心,陕西西安710119

出　　处：《陕西师范大学学报（自然科学版）》2014年第6期71-75,共5页Journal of Shaanxi Normal University：Natural Science Edition

基　　金：国家自然科学基金资助项目(81272543;81301957);中央高校基本科研业务费专项资金项目(GK201301010);陕西省自然科学基金重点项目(2014JZ005)

摘　　要：为了制备获得针对人Grn E单克隆抗体,将原核表达GST-Grn E蛋白作为免疫原,免疫Balb/c小鼠,经过融合及初步筛选,获得5株针对Grn E的单克隆细胞株.选取其中3株(1D5、2E1和3F7)制备腹水,并通过western blot和免疫荧光染色对该3株单克隆抗体进行鉴定.在此基础上,进一步对1D5和3F7两株单克隆抗体是否可以用于免疫沉淀实验进行了检测.上述实验表明,本研究成功制备了针对Grn E结构域的单克隆抗体,而且所制备的单克隆抗体具有良好的特异性.To produce monoclonal antibodies against human Grn E,GST-Grn E fusion protein was expressed using the prokaryotic expression system,then it was used to immune Balb/c mice. Finally,five hybridoma cell lines secreting Mabs against Grn E were obtained by limited clone di-lution.Three hybridomas （1D5,2E1 and 3F7）were selected to prepare ascites,then further i-dentified by the western blot and immunofluorescence staining.In addition,1D5 and 3F7 were selected for doing immunoprecipitation.The experimental results indicated that the monoclonal antibodies raised in the paper are specific against human Grn E.The success of the preparation of Mabs against Grn E provides an important tool to the study of PGRN function and its molecular mechanisms.

关 键 词：颗粒蛋白前体 单克隆抗体 Grn E 杂交瘤 

分 类 号：Q274[生物学—细胞生物学]