Abstract
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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acids / metabolism*
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Animals
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Carbon Isotopes / analysis
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Carbon Isotopes / metabolism
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Cells, Cultured / drug effects
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Cells, Cultured / metabolism*
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Chromatography, Liquid / methods
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HeLa Cells / drug effects
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HeLa Cells / metabolism
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Humans
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Indicators and Reagents
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Isotope Labeling / instrumentation
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Isotope Labeling / methods*
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Mass Spectrometry / methods*
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Mice
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Nitrogen Isotopes / analysis
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Nitrogen Isotopes / metabolism
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Protein Interaction Mapping
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Protein Processing, Post-Translational
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Proteins / metabolism*
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Swiss 3T3 Cells / drug effects
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Swiss 3T3 Cells / metabolism
Substances
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Amino Acids
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Carbon Isotopes
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Indicators and Reagents
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Nitrogen Isotopes
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Proteins