兔抗人硒蛋白P抗体的制备与鉴定  

Preparation and characterization of rabbit antibody against human selenoprotein P

作　　者：房青[1] 张明程[2] 查园园[1] 毕胜利[2] 李志伟[3] 杨建国[3] 小祝修 林晨[1] 

机构地区：[1]中国医学科学院中国协和医科大学肿瘤研究所分子肿瘤学国家重点实验室,北京100021 [2]中国疾病预防控制中心病毒病预防控制所肝炎室,北京100057 [3]北京市营养源研究所,北京100054 [4]日本东京理工大学理工学部

出　　处：《细胞与分子免疫学杂志》2004年第1期91-94,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：北京市自然科学基金资助 (No .70 0 2 0 0 7)

摘　　要：目的 :以原核表达的人硒蛋白P片段 (HSelP)制备其兔多克隆抗体并进行鉴定。方法 :在E .coli中诱导表达HSelP片段 ,经DEAEfastflow和Ni NTA Agarose柱层析纯化后 ,以其为免疫原制备兔抗HSelP抗体 ,并以Westernblot鉴定抗体的特异性。结果 :成功地表达并纯化了HSelP ,制备的兔抗HSelP抗体可有效地检测天然的SelP。结论 :以原核表达并纯化的HSelP为免疫原 ,成功地制备了兔抗该蛋白的抗体 ,Westernblot鉴定特异性良好。为进一步研究硒蛋白P的功能。AIM: To prepare rabbit antibody against human selenoprotein P (HSelP) by using HSelP expressed in the prokaryotic expression system and identify its specificity. METHODS: HSelP gene fragment was expressed in E.Coli via IPTG induction and purified through DEAE and Ni-NTA columns sequentially. The purified HSelP as immunogen was used to immunize rabbit. The specificity of rabbit anti-HSelP antibody was analyzed by Western blot. RESULTS: HSelP protein had been expressed and purified successfully. The polyclonal anti-HSelP antibody could specifically recognize natural HSelP from human serum. CONCLUSION: Polyclonal anti-HSelP antibody with high specificity and purity has been prepared by using purified HSelP as immunogen, which lays the foundation for further research on the function and distribution of HSelP, as well as developing a simple method for detecting HSelP.

关 键 词：人硒蛋白P 蛋白表达和纯化 多克隆抗体 

分 类 号：Q786[生物学—分子生物学]