The Production and Application of Ezrin Polyclonal Antibody

Abstract

Abstract: Constructed Ezrin prokaryotic expression vector pET-28a(+)-ezrin, the recombinant plasmid was transformed into E. coliBL21(DE3), induced Ezrin protein was purified through Ni-chelating affinity chromatography and purified protein was used to immunize NewZealand rabbits and Kunming mice, antiserum was removed. The titer and specificity of anti-sera were determined by ELISA, Western blottingand immunofluorescence. The specificity of the anti-Ezrin antibody was identified by Western blot and immunofluorescence. The results showedthat anti-sera antibody reacted with antigen and detected a specific band of 82 kD, and the size was agreed with the predicted molecular mass,localization dectected by inmmunofluorescence was in good agreement with the references. Finally, the anti-surem was purified by protein G.These results suggested that the Ezrin polyclonal antibody revealed high sensitivity and specificity.

Key words: Prokaryotic expression and purification , Ezrin , Animal immunization , Anti-serum , Polyclonal antibody

Zhang Hongling, Wan Jun, Yan Fei, Zhang Xinsheng, Cao Wei, Tao Wei, Huang Laiqiang. The Production and Application of Ezrin Polyclonal Antibody[J]. Biotechnology Bulletin, 2013, 0(1): 129-133.

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