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ELF3多克隆抗体的制备及其免疫定位
引用本文:汪志军,常莹,吴伟,姚津剑,何星星,林菊生,宋宇虎.ELF3多克隆抗体的制备及其免疫定位[J].华中科技大学学报(医学版),2010,39(1).
作者姓名:汪志军  常莹  吴伟  姚津剑  何星星  林菊生  宋宇虎
作者单位:1. 华中科技大学同济医学院附属同济医院肝病研究所,武汉,430030
2. 华中科技大学同济医学院附属协和医院消化内科,武汉,430022
基金项目:国家自然科学基金资助项目 
摘    要:目的 制备并鉴定兔抗小鼠胚肝血影蛋白3(embryonic liver fodrin 3,ELF3)的多克隆抗体,并探讨其在部分小鼠组织中的免疫定位.方法 合成ELF3氨基端特异性多肽片段,连接载体蛋白钥孔戚血蓝素(keyhole limpet hemocyanin,KLH),免疫新西兰大白兔,制备多克隆抗体,用ELISA方法 检测抗体效价,Western blot和免疫组织化学方法 检测抗体特异性及在小鼠组织中的表达情况.结果合成ELF3特异性多肽,并连接KLH后免疫实验兔,制备得到兔抗小鼠ELF3多克隆抗体,ELISA及Western blot检测结果显示此抗体效价及特异性高;Western blot和免疫组织化学检测结果显示ELF3在心、肝、肾组织中表达,尤其是细胞膜上表达明显.结论 本实验成功制备ELF3多克隆抗体,并证实ELF3在小鼠心、肝、肾组织细胞膜表达明显,这将为ELF3功能研究提供有力的工具.

关 键 词:胚肝血影蛋白3  抗体制备  免疫定位

Preparation of Rabbit Polyclonal Antibody against ELF3 and Its Immunolocalization in Mice
Wang Zhijun,Chang Ying,Wu Wei,Yao Jinjian,He Xianxian,Lin Jusheng,Song Xuhu.Preparation of Rabbit Polyclonal Antibody against ELF3 and Its Immunolocalization in Mice[J].Journal of Huazhong University of Science and Technology(Health Sciences),2010,39(1).
Authors:Wang Zhijun  Chang Ying  Wu Wei  Yao Jinjian  He Xianxian  Lin Jusheng  Song Xuhu
Institution:Wang Zhijun,Chang Ying,Wu Wei et al Institute of Liver Diseases,Tongji Hospital,Tongji Medical College,Huazhong University of Science , Technology,Wuhan 430030,China
Abstract:Objective To prepare and identify rabbit polyclonal antibody against embryonic liver fordrin 3(ELF3),and investigate the distribution of ELF3 in mice tissue.Methods ELF3 specific N-terminal peptide was synthesized,and conjugated to Keyhole limpet hemocyanin(KLH)as immunogen.The ELF3-KLH complex was injected into rabbits subcutaneously,and then ELF3 antibody was purified using affinity chromatography.The titer of the antibody was evaluated by ELISA.The specificity of antibody against ELF3 and immunolocalizat...
Keywords:embryonic liver fodrin 3  antibody preparation  immunolocalization
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