抗谷胱甘肽S转移酶多克隆抗体的制备及特异性鉴定

Preparation and Identification of Anti-GST Polyclonal Antibody

目的:制备谷胱甘肽S转移酶(GST)的兔多抗,并鉴定该抗体的特异性。方法:用纯化的GST标签蛋白(纯度>98%)免疫新西兰大白兔,获得GST的兔抗血清,并经HiTrap rProtein A柱纯化获得高效价高特异性的抗体;用间接ELISA法检测抗体效价,Western印迹检测抗体的特异性,并与商业化抗体进行对比。结果:通过免疫法得到了GST的兔多克隆抗体血清,抗体效价达1∶1×106,经rProtein A柱纯化后获得了高效价高特异性的抗体,其高效高特异性已达商业化抗体水平。结论:获得了GST的高效价高特异性的兔多克隆抗体。

Objective： To prepare and identify rabbit anti-glutathione S-transferase（GST） polyclonal antibody.Methods： New Zealand white rabbit was immunized by the purified GST（purity is over 98%） to produce poly clonal antiserum of GST.Then the serum were purified by HiTrap rProtein A columns to obtain high titer and spe cific antibody.Furthermore,the purified antibody was identified by indirect ELISA and Western blotting.Results： After immunization,rabbit antiserum of GST was prepared and the highest titer was 1∶1×106,its titer and specifici ty were comparable to commercial antibody.Conclusion： The results indicated that specific rabbit anti-GST poly clonal antibody with high titer was prepared.