【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、分布及建立较简便的检测方法打下了基础更多还原

【Abstract】 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.更多还原