Stable isotope labeling with amino acids in cell culture (SILAC) for studying dynamics of protein abundance and posttranslational modifications.
Stable isotope labeling with amino acids in cell culture (SILAC) is a simple and straightforward approach for in vivo incorporation of a tag into proteins for relative quantitation by mass spectrometry. SILAC is a simple, yet powerful, method for investigating the dynamics of protein abundance and posttranslational modifications. Here, we provide detailed instructions for using this method to study protein complexes, protein-protein interactions, and the dynamics of protein abundance and posttranslational modifications. We expect that SILAC will become a routine technique because of its applicability to most areas of cell biology. We have also developed a Web site (http://www.silac.org) to provide researchers with updated information about this method and related resources.
Amanchy
Ramars
R
McKusick-Nathans Institute of Genetic Medicine and Department of Biological Chemistry and Oncology, Johns Hopkins University, Baltimore, MD 21218, USA.
Kalume
Dario Eluan
DE
Pandey
Akhilesh
A
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D002247:Carbon Isotopes / Q000032:analysis / Q000378:metabolism; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods; D006367:HeLa Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007202:Indicators and Reagents; D007553:Isotope Labeling / Q000295:instrumentation / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D009587:Nitrogen Isotopes / Q000032:analysis / Q000378:metabolism; D025941:Protein Interaction Mapping; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism*; D041701:Swiss 3T3 Cells / Q000187:drug effects / Q000378:metabolism
D000596:Amino Acids; D002247:Carbon Isotopes; D007202:Indicators and Reagents; D009587:Nitrogen Isotopes; D011506:Proteins