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Updated: Mar 19, 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

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Identification of Associated Proteins by Immunoprecipitation and Mass Spectrometry Analysis

Xiumei Cao1, Jianshe Yan2

  • 1Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, South Chongqing Road No. 280, Shanghai, 200025, China.

Methods in Molecular Biology (Clifton, N.J.)
|June 9, 2016
PubMed

Protein-protein interactions play central roles in intercellular and intracellular signal transduction. Impairment of protein-protein interactions causes many diseases such as cancer, cardiomyopathies, diabetes, microbial infections, and genetic and neurodegenerative disorders. Immunoprecipitation is a technique in which a target protein of interest bound by an antibody is used to pull down the protein complex out of cell lysates, which can be identified by mass spectrometry. Here, we describe the protocol to immunoprecipitate and identify the components of the protein complexes of ElmoE in Dictyostelium discoideum cells.

Keywords:
Dictyostelium discoideumImmunoprecipitationMass spectrometryProtein-protein interaction

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