Identification of interaction between ISCA1 and NLS-RARα by co-immunoprecipitation in mammalian cells

AIM:To verify the interaction between ISCA1 and NLS-RARα by co-immunoprecipitation and Western blotting.METHODS:The eukaryotic expression vector pCMV-HA-NLS-RARα and pCMV-Myc-ISCA1 were constructed,identified and then transfected into human embryo kidney 293 cells.Co-immunoprecipitation and Western blotting were used to investigate the interaction between ISCA1 and NLS-RARα.RESULTS:After being verified,eukaryotic expression vectors were co-transfected into HEK 293 cells,HA-NLS-RARα protein was then immunoprecipitated by anti-HA polyclonal antibody,and Myc-ISCA1 protein was detected by western blotting with anti-Myc monoclonal antibody from the immunoprecipitared complex.CONCLUSION:Eukaryotic expression vectors with HA-NLS-RARα and Myc-ISCA1 are constructed successfully.The interaction between ISCA1 and NLS-RARα is identified by co-immunoprecipitation and Western blotting in vitro.