Dundee Cell Products
SILAC Tissue Culture Media
Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC) is a metabolic labelling technique for MS-based quantitative proteomics. Differentially labelled samples are mixed early in the experimental process and analyzed together. The peptide peaks of the differentially labelled samples can accurately quantified relative to each other and used to determine the peptide and protein ratios. Processing and measuring the samples together ensures accurate reproducibility and quantitation.
SILAC labelling requires the cells comprising each sample to be grown in isotopically different tissue culture media. For traditional SILAC, the molecular masses of either the lysine and/or arginine within the media yield media types more commonly referred to as "light", "medium" and "heavy" media. The differences in molecular mass are achieved by the differential use of heavy carbon (13C) and nitrogen (15N) isotopes. This labelling, combined with tryptic digestion of the sample, results in the proteolytic cleavage of proteins at arginine and lysine, thus ensuring that every peptide within the mixture is isotopically labelled.
Dundee Cell Products were the first company in the world to produce ready-to-use SILAC media commercially. Using ready-to-use media saves time and abolishes the risk of contamination; all you need do is add dialyzed serum. DCP produce a wide range of SILAC media , we are also able to produce SILAC media to your specific requirements.
The labelled amino acids use to produce our media have a purity > 99%. The standard means of abbreviating the different amino acid isotopes used within SILAC media are as follows:
R6 = L-Arginine-13C6 HCl
R10 = L-Arginine-13C6, 15N4 HCl
K4 = L-Lysine-4,4,5,5-D4 HCl
K6 = L-Lysine-13C6, HCl
K8 = L-Lysine-13C6,15N2 HCl
Together these are combined to make the different types of media:
R0K0 = unlabelled/ light label
R6K4 = medium label*
R6K6 = medium label*
R10K8 = heavy label
*The type of medium media used depends on how many samples are to be pooled.
Dundee Cell Products also supply a range of SILAC reagents that includes dialyzed foetal bovine and horse serum for SILAC.