Protein Interaction Analysis in HEK-293 Cells Using Co-Immunoprecipitation

January 05, 2024

Immunoprecipitation with an Anti-Epitope Tag Affinity Gel to Study Protein-Protein Interactions

01:52 min

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Cell Lysis of Transfected HEK293 Cells and Protein Sample Preparation for Immunoprecipitation

01:14 min

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Preclearing of Transfected HEK293 Protein with the Protein G Gel and Capturing Protein Complexes Using Epitope Tag Affinity Gel

01:17 min

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Eluting Epitope Tag Precipitated Proteins and Immunoblotting for Protein-Protein Interaction Studies

02:58 min

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Hiroko Shinjo, Gaku Nagano, Shogo Ishii, Natsumi Himeno, Yutaro Yamamoto, Junji Sagawa, Ryuta Baba, Gentaro Egusa, Noboru Hattori, Haruya Ohno

¹Department of Molecular and Internal Medicine, Graduate School of Biomedical & Health Sciences,Hiroshima University

Summary

Protein-protein interactions are important for elucidating the function of target proteins, and co-immunoprecipitation (co-IP) can easily confirm PPIs. We transiently transfected a plasmid encoding an epitope-tagged protein into HEK-293 cells and developed an immunoprecipitation method to easily confirm the binding of two target proteins.