2011•Biaoji mianyi fenxi yu linchuangRequires access
Preparation of Polyclonal Antibody Against Human CPNE5
Ming Fan
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Abstract
To prepare polyclonal antibody against human CPNE5,the 423bp specific sequence of CPNE5(626~1048bp) was cloned into the vector pGEX-5X-1.The recombinant plasmid was transformed into host strain E.coli BL21 and induced by IPTG.Recombinant protein was purified by SDS-PAGE and detected by mass spectrum.The protein was then used as antigen to immunize rabbit by the conventional method.The polyclonal antibody was purified by protein A sepharose CL-4B.The results showed that the antigen was successfully expressed and purified.The high affinity of the antiserum was confirmed by ELISA.The results of Western blot showed this antibody could specifically recognize CPNE5 protein.The CPNE5 antibody with high affinity and specificity was successfully prepared.