title: Multiplex Sequential Immunoprecipitation of Insulin Secretory Granule Proteins from Radiolabeled Pancreatic Islets.
abstract: Pulse radiolabeling of cells with radioactive amino acids is a common method for tracking the biosynthesis of proteins. Specific proteins can then be immunoprecipitated and analyzed by electrophoresis and imaging techniques. This chapter presents a protocol for the biosynthetic labeling of pancreatic islets with35S-methionine, followed by multiplex sequential immunoprecipitation of insulin and three other secretory granule accessory proteins. This provided a means of distinguishing those pancreatic islet proteins with different biosynthetic rates in response to the media glucose concentrations.
authors.affiliation: Laboratory of Neuroproteomics, Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Rua Monteiro Lobato 255 F/01, Cidade Universitária Zeferino Vaz, 13083-862, Campinas, Brazil.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Guest
mesh_terms: D000818:Animals; D004586:Electrophoresis / Q000379:methods; D047468:Immunoprecipitation / Q000379:methods*; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000378:metabolism*; D007553:Isotope Labeling; D051381:Rats; D022142:Secretory Vesicles / Q000378:metabolism*
chemical_list: D007328:Insulin
keywords: Electrophoresis; Immunoprecipitation; Insulin; Pancreatic islets; Pulse radiolabeling; Secretory granule proteins