Author Spotlight: Unraveling the Molecular Mechanisms of Brown and Beige Adipocyte Regulation

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

doi:10.3791/66085

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

January 5th, 2024

Author Spotlight: Unraveling the Molecular Mechanisms of Brown and Beige Adipocyte Regulation

01:52 min

•

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

01:14 min

•

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

01:17 min

•

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

02:58 min

•

Hiroko Shinjo¹, Gaku Nagano¹, Shogo Ishii¹, Natsumi Himeno¹, Yutaro Yamamoto¹, Junji Sagawa¹, Ryuta Baba¹, Gentaro Egusa¹, Noboru Hattori¹, Haruya Ohno¹

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

Summary
Automatic Translation

العربية (Arabic)
中文 (Chinese)
dansk (Danish)
Nederlands (Dutch)
français (French)
Deutsch (German)
עברית (Hebrew)
हिंदी (Hindi)
italiano (Italian)
日本語 (Japanese)
한국어 (Korean)
norsk (Norwegian)
português (Portugese)
русский (Russian)
español (Spanish)
svenska (Swedish)
Türkçe (Turkish)

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.

Article

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

Get cutting-edge science videos from JoVE sent straight to your inbox every month.