Matriptase-2多克隆抗体的制备及鉴定.pdf

472 Chin J Hemorh.2014;24(4) doi:10.3969/j.issn.1009-881X.2014.04.002 Matriptase-2多克隆抗体的制备及鉴定 赟 1. 215123 2. 215006 Matriptase-2 Matriptase-2Glu81-Gly228 摘要：目的 方法 pMAL-C2X BL21(DE3)IPTG Matriptase-2protein G 结果 Matriptase-2Western blot Matriptase-2Matriptase-2 结论 关键词Matriptase-2 中图法分类号R55 文献标识码A 文章编号1009-881X(2014)04-0472-04 Generation and Identifi cation of a Polyclonal Antibody to Matripase-2 1 2 1 1,2 YANG Jian-feng , ZHAO Yun-xiao , ZUO Bin , HE Yang (1.Cyrus Tang Hematology Center, Soochow University, Suzhou, Jiangsu, 215123, China; 2.The First Affi liated Hospital of Soochow University, Jiangsu Institute of Hematology, Suzhou, Jiangsu, 215006, China) Abstract Objective To investigate the expression of Matriptase-2 recombinant protein in E.coli and the generation of its polyclonal antibody. Methods Extracellular region (Glu81-Gly228) proximal to transmembrane domain of Matriptase-2 was cloned into pMAL-C2X. The recombinant plasmid was transformed to BL21 (DE3) bacteria. After IPTG induction, the total protein was extracted and the recombinant Matriptase-2 protein was purifi ed with amylose resin. The Matriptase-2 recombinant protein was used to immunize New Zealand Rabbits to prepare serum with anti-Matriptase-2 polyclonal antibody. The antibody was purifi ed from serum by protein G sepharose 4B. Results Recombinant Matriptase-2 protein was successfully expressed in E.coli and anti-serum with high titer was obtained. Western blotting result showed that the antibody specifi cally recognized Matriptase-2. Conclusion It is