title: Stable isotope labeling of mammals (SILAM) for in vivo quantitative proteomic analysis.
abstract: Metabolic labeling of rodent proteins with ¹⁵N, a heavy stable isotope of nitrogen, provides an efficient way for relative quantitation of differentially expressed proteins. Here we describe a protocol for metabolic labeling of rats with an ¹⁵N-enriched spirulina diet. As a case study, we also demonstrate the application of ¹⁵N-enriched tissue as a common internal standard in quantitative analysis of differentially expressed proteins in neurodevelopment in rats at two different time points, postnatal day 1 and 45. We briefly discuss the bioinformatics tools, ProLucid and Census, which can easily be used in a sequential manner to identify and quantitate relative protein levels on a proteomic scale.
authors.affiliation: Department of Chemical Physiology, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037, USA.
authors.forename: Navin
authors.initials: N
authors.lastname: Rauniyar
authors.forename: Daniel B
authors.initials: DB
authors.lastname: McClatchy
authors.forename: John R
authors.initials: JR
authors.lastname: Yates
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000254:growth & development / Q000378:metabolism*; D001923:Brain Chemistry; D004032:Diet; D005260:Female; D018507:Gene Expression Regulation, Developmental; D007553:Isotope Labeling; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D009587:Nitrogen Isotopes; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D051381:Rats; D053545:Spirulina / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D009419:Nerve Tissue Proteins; D009587:Nitrogen Isotopes; D010446:Peptide Fragments