Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) – an Introduction for Biologists
Robert L.J. Graham,
Michael J. Sweredoski,
Since its introduction in 2002 ‘stable isotope labeling by amino acids in cell culture’ (SILAC) has become a major tool for quantitative proteomics. Stable isotopes are incorporated into proteins by introduction of labeled amino acids in growth medium. This methodologys compatibility with virtually all cell culture conditions, ease of implementation into existing workflows and the high quality quantitative data that can be obtained have made it the quantitative method of choice for many laboratories. Although originally used for mammalian cell cultures, it has been adapted to a number of organisms including yeast, bacteria, plants and higher organisms. This review will look at the use of SILAC as a key tool in quantitative biology. The purpose of this review is to furnish the reader with a basic understanding of the fundamental principles of SILAC including its implementation, application, bioinformatic tools for its analyses and potential problems that one should be mindful of.
Keywords: Stable isotope labeling by amino acids in cell culture, SILAC, proteomics, mass spectrometry, Enzymatic, Peptides (iTRAQ), Proteolytic Peptides, Saccharomyces Cerevisiae, Deuterium, Thiol-Specific, Tri-Deuterium, Escherichia Coli, LC-MS/MS, MALDI-MS, Phosphorylation
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