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Updated: Apr 26, 2026

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation
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A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

Published on: September 24, 2012

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Identification and validation of protein-protein interactions by combining co-immunoprecipitation, antigen

Frederik Sommer1, Timo Mühlhaus, Dorothea Hemme

  • 1Molecular Biotechnology and Systems Biology, TU Kaiserslautern, Paul-Ehrlich-Str. 23, 67663, Kaiserslautern, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|July 26, 2014
PubMed

Co-immunoprecipitation (coIP) in combination with mass spectrometry (MS) is a powerful tool to identify potential protein-protein interactions. However, unspecifically precipitated proteins usually result in large numbers of false-positive identifications. Here we describe a detailed protocol particularly useful in plant sciences that is based on (15)N stable isotope labeling of cells, (14)N antigen titration, and coIP/MS to distinguish true from false protein-protein interactions.

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