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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
OBJECTIVE: To prepare and identify rabbit polyclonal antibody by using prokaryotic expression fragment of human selenoprotein P (HSelP). Methods: The HSelP fragment was induced in E.coli. After purified by DEAEfastflow and Ni NTA Agarose column chromatography, the rabbit anti-HSelP antibody was prepared as an immunogen. The specificity of the antibody was identified by Western blot. Results: HSelP was successfully expressed and purified. The prepared rabbit anti-HSelP antibody could effectively detect native SelP. CONCLUSION: The rabbit anti-rabbit antibody was successfully prepared using the prokaryotic expressed and purified HSelP as an immunogen. The specificity of Western blot was good. This laid the foundation for further study on the function and distribution of selenoprotein P and the establishment of a more convenient detection method