title: Biochemical purification of native immune protein complexes.
abstract: Protein complex purification represents a powerful approach to identify novel players in plant innate immunity. However, the identification of interacting protein partners within a natural context has been a challenge for researchers. In this chapter, we describe a method of immunoaffinity chromatography using purified, antibodies to isolate native protein complexes from wild-type tissue. We detail the antibody purification and immobilization steps in addition to the co-immunoprecipitation protocol. In addition, a method to prepare protein samples for mass spectroscopy analysis is described. This straightforward protocol has been used to isolate and identify novel components of Arabidopsis immunity-associated protein complexes.
authors.affiliation: Department of Plant Pathology, University of California, Davis, CA, USA.
authors.forename: James M
authors.initials: JM
authors.lastname: Elmore
authors.forename: Gitta
authors.initials: G
authors.lastname: Coaker
mesh_terms: D000936:Antigen-Antibody Complex / Q000302:isolation & purification*; D017360:Arabidopsis / Q000737:chemistry / Q000276:immunology; D029681:Arabidopsis Proteins / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D047468:Immunoprecipitation / Q000295:instrumentation / Q000379:methods*; D057865:Plant Immunity / Q000276:immunology
chemical_list: D000936:Antigen-Antibody Complex; D029681:Arabidopsis Proteins