A practical recipe for stable isotope labeling by amino acids in cell culture (SILAC)

By Shao-En Ong, Matthias Mann. Nature Protocols, 2006

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Multiplex peptide stable isotope dimethyl labeling for quantitative proteomics (2009, background): “A popular protocol for isotopic labeling of proteins in eukaryotic cells is SILAC, where cells are grown in culture media lacking essential (auxotrophic) amino acid(s”
Amyloid-like aggregates sequester numerous metastable proteins with essential cellular functions. (2011, methodology): “To test this hypothesis, we performed a sensitive, quantitative proteomic analysis of the b protein interactome using SILAC (stable isotope labeling with amino acids in cell culture) (Ong and Mann, 2006) and peptide identification by tandem mass spectrometry (LC-MS/MS).”
SILAC mouse for quantitative proteomics uncovers kindlin-3 as an essential factor for red blood cell function. (2008, methodology): “In-solution digestion of proteins was performed as described by Ong and Mann (2006).”
In-gel digestion for mass spectrometric characterization of proteins and proteomes (2006, methodology): “or by mixing SILAC-labeled protein mixtures before separatio”