Co-immunoprecipitation and semi-quantitative immunoblotting for the analysis of protein-protein interactions

STAR Protoc. 2021 Jul 6;2(3):100644. doi: 10.1016/j.xpro.2021.100644. eCollection 2021 Sep 17.

Abstract

Co-immunoprecipitation (co-IP) of protein complexes from cell lysates is widely used to study protein-protein interactions. However, establishing robust co-IP assays often involves considerable optimization. Moreover, co-IP results are frequently presented in non-quantitative ways. This protocol presents an optimized co-IP workflow with an analysis based on semi-quantitative immunoblot densitometry to increase reliability and reproducibility. For complete details on the use and execution of this protocol, please refer to Burckhardt et al. (2021).

Keywords: Cell Biology; Gene Expression; In Situ Hybridization; Microscopy; Model Organisms; Molecular Biology; Single Cell; Single-molecule Assays.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Electrophoresis / methods
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Immunoblotting / methods*
  • Immunoprecipitation / methods*
  • Kidney / cytology
  • Kidney / metabolism
  • Protein Interaction Maps / physiology*
  • Proteomics / methods
  • Transfection

Substances

  • Green Fluorescent Proteins