Tianjin Med J ›› 2016, Vol. 44 ›› Issue (10): 1195-1198.doi: 10.11958/20160317

• Cell and Molecular Biology • Previous Articles     Next Articles

The preparation of the mouse monoclonal antibodies specific for the DNAH2 protein

CHANG Lixian, ZHANG Jingliao, REN Yuanyuan, SUN Congcong, WAN Yang, AN Wenbin, ZHANG Yingchi, YUAN Weiping, ZHU Xiaofan   

  1. State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, China
  • Received:2016-04-19 Revised:2016-07-08 Published:2016-10-15 Online:2016-10-21
  • Contact: ZHU Xiaofan E-mail:clx.886@163.com

Abstract: Objective To prepare specific mouse monoclonal antibodies against Homo sapiens dynein, axonemal, heavy chain 2 (DNAH2). Methods Firstly, recombinant plasmid encoding His tagged immunogen, targeting N-terminal sequence of DNAH2 protein (1- 300 aa), in E. coli was constructed. IPTG was used to induce the expression of Hisimmunogen, which was then purified and immunized in BALB/c mice. Hybridoma cells were obtained through the fusion between myeloma cells and splenocytes isolated from BALB/c mice. Finally, ELISA and Western blot assays were performed to screen the positive hybridoma. Results IPTG was used efficiently to induce the expression of DNAH2 immunogen in E. coli. DNAH2 protein bands were detected in screened positive hybridoma. Conclusion Mouse monoclonal antiDNAH2 antibody is prepared successfully.

Key words: antibodies, monoclonal, hybridomas, plasmids, recombination, genetic, DNAH2