title: Identification and validation of protein-protein interactions by combining co-immunoprecipitation, antigen competition, and stable isotope labeling.
abstract: 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.
authors: Molecular Biotechnology and Systems Biology, TU Kaiserslautern, Paul-Ehrlich-Str. 23, 67663, Kaiserslautern, Germany.
authors: Frederik
authors: F
authors: Sommer
authors: Timo
authors: T
authors: Mühlhaus
authors: Dorothea
authors: D
authors: Hemme
authors: Daniel
authors: D
authors: Veyel
authors: Michael
authors: M
authors: Schroda
mesh_terms: D053000:Analytic Sample Preparation Methods; D000906:Antibodies / Q000276:immunology; D000941:Antigens / Q000276:immunology*; D001667:Binding, Competitive; D018929:Cell Culture Techniques; D002696:Chlamydomonas / Q000166:cytology; D047468:Immunoprecipitation / Q000379:methods*; D007553:Isotope Labeling / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D009587:Nitrogen Isotopes / Q000737:chemistry; D025941:Protein Interaction Mapping / Q000379:methods*
chemical_list: D000906:Antibodies; D000941:Antigens; D009587:Nitrogen Isotopes