title: [Improvement of immunoprecipitation/immunoblotting of endogenous protein by TrueBlot antibody].
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.
  
  
  CONCLUSIONS
  TrueBlot antibody prefers the immunoblot antibody to immunoprecipitation antibody, and decreases the interruption of immunoprecipitation antibody to display clear protein band.
authors: Department of Clinical Laboratory, Xiangya School of Medicine, Central South University, Changsha 410013, China huangjianjunhn@hotmail.com.
authors: Jianjun
authors: J
authors: Huang
authors: Keqian
authors: K
authors: Xu
authors: Jianxin
authors: J
authors: Luo
authors: Jianxiong
authors: J
authors: Peng
mesh_terms: D000906:Antibodies; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D011506:Proteins; D014162:Transfection
chemical_list: D000906:Antibodies; D011506:Proteins