Stable isotope labeling with amino acids in cell culture (SILAC), has recently gained popularity for its ability to compare the expression levels of hundreds of proteins in a single experiment. SILAC was developed at the Center for Experimental BioInformatics (CEBI)1 as a simple and accurate approach for MS-based quantitative proteomics. The method relies on the incorporation of unlabeled and D, 13C, and/or 15N-labeled amino acids added to the growth media of separately cultured cell lines, giving rise to cells containing either “light” or “heavy” proteins, respectively.
Upon mixing lysates collected from these cells, digests of the protein populations are analyzed by mass spectometry. Using this method, one can compare the expression levels for hundreds of proteins. This valuable and robust proteomics tool is being used in many different applications, including:
Sigma now offers two new media for SILAC applications. These new media are deficient in arginine, leucine, and lysine and are designed to be supplemented with istopically labeled versions of those amino acids.
References 1. Ong S.E. et al. (2002). Mol Cell Proteomics 1, 376–86. 2. Gronborg M. et al. (2006). Mol Cell Proteomics 5, 157–171. 3. Milner E., Barnea E., Beer I, and Admon, A. (2006). Mol Cell Proteomics 5: 357–365. 4. de Godoy LM, Olsen JV, de Souza GA, Li G, Mortensen P., Mann M. (2006). Genome Biol. 7(6): R50.
RPMI-1640 Medium With L-glutamine and sodium bicarbonate. Without arginine, leucine, lysine, and phenol red, liquid, sterile-filtered, cell culture tested
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