Identification of interaction between ISCA1 and NLS-RARα by co-immuno precipiration 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-immu-noprecipitation and Western blotting were used to investigate the interaction between ISCA1 and NLS-RARα. RESULTS: After being verified, eukaryotic expression rectors were co-transfected into HEK 293 cells, HA-NLS-RARα protein was then immunoprecipi-tated 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.
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万方数据
维普
