ELF3多克隆抗体的制备及其免疫定位  

Preparation of Rabbit Polyclonal Antibody against ELF3 and Its Immunolocalization in Mice

作　　者：汪志军[1] 常莹[1] 吴伟[1] 姚津剑[1] 何星星[1] 林菊生[1] 宋宇虎[2] 

机构地区：[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4300302 [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

出　　处：《华中科技大学学报（医学版）》2010年第1期6-8,17,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30600277)

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 hemo-cyanin,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。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 immunolocalization of ELF3 were evaluated by using Western blot and immunohistochemistry. Results Rabbit polyclonal antibody against ELF3 was prepared by the immunization of ELF3 KLH complex. ELISA and Western blot results showed the antibody against ELF3 had high titer and specificity. Western blot and immunohistochemical studies demonstrated ELF3 was expressed in the mouse heart, liver, brain and kidney tissue, particular on the cell membrane. Conclusion The preparation of polyclonal antibody against ELF3 was successful due to its high titer and specificity; ELF3 was expressed in the mice heart,liver, and kidney, particular on the cell membrane. It will provide an excellent tool for further study on the ELF3 function.

关 键 词：胚肝血影蛋白3 抗体制备 免疫定位 

分 类 号：R392.11[医药卫生—免疫学]