Tuesday, November 16, 2010
SILAC: Stable Isotope Labeling with Amino acid in Cell culture
SILAC is a simple and straight forward approach for in vivo incorporation of a label into proteins for MS-based quantitative proteomics. SILAC relies on metabolic incorporation of a given "light" or "heavy" form of the amino acid into proteins. The method relies on the incorporation of amino acids with substituted stable isotopic nuclei (e.g. deuterium, 13C, 15N). This in an experiment, two cell populations are grown in cultre media that are identical except that one of them contains a "light" and the other a "heavy" form of a particular amino acid (e.g. 12C and 13C labeled L-lysine, respectively).When the labeled analog of an amino acid is supplied to cells in culture instead of the natural amino acid, it is incorporated into all synthesized proteins. After a number of cell division, each instance of this particular amino acid will be replaced by its isotope labeled analog. Since there is hardly any chemical difference between the labeled amino acid and the natural amino acid isotopes, the cells behave exactly like the control cell population grown in the presence of normal amino acid. It is efficient and reproducible as the incorporation of the isotope is 100%.We anticipate of SIALC will lead to is use as a routine technology in all areas of cell biology.
The above is from http://silac.org/index_html