The preparation of the mouse monoclonal antibodies specific for the DNAH2 protein
10.11958/20160317
- VernacularTitle:针对人DNAH2蛋白的鼠源单克隆抗体的制备
- Author:
Lixian CHANG
;
Jingliao ZHANG
;
Yuanyuan REN
;
Congcong SUN
;
Yang WAN
;
Wenbin AN
;
Yingchi ZHANG
;
Weiping YUAN
;
Xiaofan ZHU
- Publication Type:Journal Article
- Keywords:
antibodies,monoclonal;
hybridomas;
plasmids;
recombination,genetic;
DNAH2
- From:
Tianjin Medical Journal
2016;44(10):1195-1198
- CountryChina
- Language:Chinese
-
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 His-immunogen, 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 anti-DNAH2 antibody is prepared successfully.
