title: Co-immunoprecipitation for Deciphering Protein Interactomes.
abstract: A single protein is often capable of binding with many partners, enabling potential effects on either protein, such as modifying its expression or activity. However, due to the complex nature of in vivo systems, it is often difficult to perform nontargeted assays with a protein of interest. Methods in discovery proteomics must be used to find potential interactors to pave the way for additional, more focused studies. This protocol describes the biological steps needed to create an interactome focused on a single protein target through co-immunoprecipitation.
authors.affiliation: Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Rua Monteiro Lobato, 255, 13083-862, Campinas, SP, Brazil.
authors.forename: Bradley J
authors.initials: BJ
authors.lastname: Smith
authors.affiliation: Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Rua Monteiro Lobato, 255, 13083-862, Campinas, SP, Brazil.
authors.forename: Juliana S
authors.initials: JS
authors.lastname: Cassoli
authors.affiliation: Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Rua Monteiro Lobato, 255, 13083-862, Campinas, SP, Brazil.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Guest
authors.affiliation: Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Rua Monteiro Lobato, 255, 13083-862, Campinas, SP, Brazil. dmsouza@unicamp.br.|UNICAMP's Neurobiology Center, Campinas, SP, Brazil. dmsouza@unicamp.br.
authors.forename: Daniel
authors.initials: D
authors.lastname: Martins-de-Souza
mesh_terms: D002458:Cell Fractionation / Q000379:methods; D002846:Chromatography, Affinity / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D007202:Indicators and Reagents; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D040901:Proteomics / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D007202:Indicators and Reagents
keywords: Co-immunoprecipitation; Immunoaffinity purification; Interactome; Protein-protein interactions; Shotgun proteomics