Scientific Publications

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

Publication TypeJournal Article
AuthorsOng, SE, and Mann M.
AbstractStable isotope labeling by amino acids in cell culture (SILAC) is a simple, robust, yet powerful approach in mass spectrometry (MS)-based quantitative proteomics. SILAC labels cellular proteomes through normal metabolic processes, incorporating non-radioactive, stable isotope-containing amino acids in newly synthesized proteins. Growth medium is prepared where natural ("light") amino acids are replaced by "heavy" SILAC amino acids. Cells grown in this medium incorporate the heavy amino acids after five cell doublings and SILAC amino acids have no effect on cell morphology or growth rates. When light and heavy cell populations are mixed, they remain distinguishable by MS, and protein abundances are determined from the relative MS signal intensities. SILAC provides accurate relative quantification without any chemical derivatization or manipulation and enables development of elegant functional assays in proteomics. In this protocol, we describe how to apply SILAC and the use of nano-scale liquid chromatography coupled to electrospray ionization mass spectrometry for protein identification and quantification. This procedure can be completed in 8 days.
Year of Publication2012
JournalNature protocols
Volume1
Issue6
Pages2650-60
Date Published (YYYY/MM/DD)2012/01/06
ISSN Number1754-2189
DOI10.1038/nprot.2006.427
PubMedhttp://www.ncbi.nlm.nih.gov/pubmed/17406521?dopt=Abstract