Improvement of immunoprecipitation/immunoblotting of endogenous protein by TrueBlot antibody
10.11817/j.issn.1672-7347.2014.07.007
- VernacularTitle:利用TrueBlot抗体改善内源性蛋白的免疫沉淀/免疫印迹分析
- Author:
Jianjun HUANG
;
Keqian XU
;
Jianxin LUO
;
Jianxiong PENG
- Publication Type:Journal Article
- Keywords:
endogenous protein;
immunoprecipitation;
immunoblotting;
TrueBlot antibody
- From:
Journal of Central South University(Medical Sciences)
2014;(7):694-697
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To improve the immunoblotting of immunoprecipitated proteins and decrease the interference of immunoprecipitation antibody in the interaction of endogenous proteins. Methods:Transient transfect cells with fusion protein expression vector containing the targeted S5b gene and the FLAG tag, the transfected cells or untransfected cells were harvested to study the exogenous or endogenous protein interaction. The total cell lysate was immunoprecipitated by specific antibody. Then the eluted immunocomplex was separated by SDS-PAGE, and the TrueBlot antibody or conventional antibody was used as the secondary antibody for immunoblotting detection of S5b and its partner (Rpt1 and Rpt2). Results:Clear immunoblotting bands for S5b, Rpt1 and Rpt2 were obtained. Conclusion:TrueBlot antibody prefers the immunoblot antibody to immunoprecipitation antibody, and decreases the interruption of immunoprecipitation antibody to display clear protein band.
