Analysis of protein–protein interaction by Co-IP in human cells

Zhenyuan Tang, Yoshinori Takahashi

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

While there are various approaches available to analyze protein–protein interactions, coimmunoprecipitation (co-IP) remains one of the most classic and commonly used methods to discover novel protein interactions or to determine the physical association of proteins. The assay begins with the preparation of total cell or tissue lysate in an appropriate lysis buffer. Protein of interest in the lysate is captured using a specific antibody and precipitated along with its binding proteins using a resin. After a series of washes to remove nonbound proteins in the lysate, the resultant immune complexes are subjected to immunoblotting, in-gel protein staining, or mass spectrometry to determine the protein–protein interaction of interest. In this chapter, a standard IP/co-IP protocol is described and potential problems and troubleshooting are discussed.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages289-296
Number of pages8
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1794
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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