title: Sequential Immunoprecipitation of Secretory Vesicle Proteins from Biosynthetically Labelled Cells.
abstract: Pulse radiolabelling of cells with radioactive amino acids is a common method for studying the biosynthesis of proteins. The labelled proteins can then be immunoprecipitated and analysed by electrophoresis and imaging techniques. This chapter presents a protocol for the biosynthetic labelling and immunoprecipitation of pancreatic islet proteins which are known to be affected in psychiatric disorders such as schizophrenia.
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. paulcguest@yahoo.com.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Guest
mesh_terms: D000918:Antibody Specificity; D002847:Chromatography, Agarose / Q000379:methods; D004586:Electrophoresis / Q000379:methods; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D047468:Immunoprecipitation / Q000295:instrumentation / Q000379:methods*; D007164:Immunosorbents; D007328:Insulin / Q000032:analysis* / Q000096:biosynthesis; D007515:Islets of Langerhans / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods; D008715:Methionine / Q000032:analysis; D045706:Proprotein Convertase 2 / Q000032:analysis* / Q000096:biosynthesis; D022142:Secretory Vesicles / Q000737:chemistry* / Q000201:enzymology; D013462:Sulfur Radioisotopes / Q000032:analysis; D014508:Urea
chemical_list: D007164:Immunosorbents; D007328:Insulin; D013462:Sulfur Radioisotopes; D014508:Urea; D008715:Methionine; C581329:PCSK2 protein, human; D045706:Proprotein Convertase 2; D005947:Glucose
keywords: Electrophoresis; Immunoprecipitation; Insulin; Pancreatic islets; Pulse-chase radiolabelling; Secretory granule proteins