抗nestin多克隆抗体的制备
閸忔娊鏁拠宥忕窗nestin基因,,nestin基因;融合表达;,抗体制备,1材料和方法,2结果,3讨论,【参考文献】
Preparation of polyclonal antibody to nestin
LI JunLin1,HAN Hua1,LAN Li2,YANG Xi1,LI JunFeng1,WANG YaRong3,ZHU YangTing1
1Department of Medical Genetics & Development Biology,School of Basic Medicine,Fourth Military Medical University,Xian 710033,2Department of Pediatrics,3Department of Radiology,Tangdu Hospital,Fourth Military Medical University,Xian 710038,China
【Abstract】AIM: To clone and express the Cfragment of cDNA encoding mouse nestin peptide and to obtain specific polyclonal antibody.METHODS:The Cfragment of cDNA encoding mouse nestin peptide was obtained with the method of RTPCR.The correctly identified sequence was cloned into the prokaryotic expression vector pET32a to construct pET32anestinC expression vector.6×HisnestinC fusion protein was expressed in E.coli BL21 with IPTG induction.After purification,the fusion protein was used to immunize New Zealand rabbits to get the rabbit antinestin serum.The specificity of the ployclonal antinestin antibody was analyzed by Western Blotting and immunohistochemical methods.RESULTS: The ployclonal antinestin antibody was successfully obtained by immunizing New Zealand rabbits with 6×HisnestinC fusion protein.The specificity of antibody was confirmed by Western Blotting and immunohistochemical methods.CONCLUSION:pET32 anestinC expression vector is constructed and highly expressed in BL21. ......
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