title: Affinity Purification of Protein Complexes Using TAP Tags.
abstract: This protocol is used for the isolation and analysis of protein complexes using the tandem affinity purification (TAP) tag system. The protocol describes the purification of a protein fused to a TAP tag comprised of two protein A domains and the calmodulin binding peptide separated by a TEV cleavage site. This is a powerful technique for rapid purification of protein complexes and the analysis of their stoichiometric composition, posttranslational modifications, structure, and functional activities.
authors.affiliation: Department of Cell Biology, Howard Hughes Medical Institute, Harvard Medical School, Boston, MA, USA.
authors.forename: Erica
authors.initials: E
authors.lastname: Gerace
authors.affiliation: Department of Cell Biology, Howard Hughes Medical Institute, Harvard Medical School, Boston, MA, USA. Electronic address: danesh@hms.harvard.edu.
authors.forename: Danesh
authors.initials: D
authors.lastname: Moazed
mesh_terms: D000818:Animals; D002147:Calmodulin / Q000737:chemistry; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D011506:Proteins / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae; D012568:Schizosaccharomyces; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D002147:Calmodulin; D007074:Immunoglobulin G; D011506:Proteins
keywords: Affinity purification; Calmodulin binding (CAM-B); Calmodulin elution (CAM-E); IgG-Sepharose; Lysis buffer; Protease cleavage; Protein lysates; TAP tags; TEV cleavage (TEV-C)