SILAC Reagents and Kits

The Thermo Scientific Pierce SILAC Protein Quantitation Kits with DMEM or RPMI 1640 contain all reagents necessary for successful isotope metabolic protein labeling, enabling quantitation of protein expression levels from differentially treated cell populations. Stable isotope labeling with amino acids in cell culture (SILAC) is a simple and accurate method to quantify differential changes in the proteome. SILAC uses metabolic incorporation of nonradioactive ¹³C- or ¹⁵N-labeled amino acids, referred to as “heavy” amino acids, into proteins using specially formulated media, either DMEM or RPMI 1640. Typical experiments involve growing two cell populations that are identical except that one contains the natural amino acid, referred to as “light,” and the other contains the heavy form (e.g., ¹²C₆ and ¹³C₆ L-lysine, respectively). Heavy ¹³C₆ ¹⁵N₄ L-Arginine, available separately (Product No. 89990), is often added to enhance peptide isotope label coverage. Protein levels in one sample are then altered through chemical treatment or genetic manipulation. Equal concentrations of cell lysate from both cell populations are combined for sample processing and subsequent protein separation by SDS-PAGE. Proteins are digested with trypsin to generate peptides for mass spectrometry (MS) and quantitation of isotopic peptide pairs When combined with Thermo Scientific Protein/Peptide Sample Enrichment Products, the Pierce SILAC Kits allow MS identification and quantitation of low-abundance proteins, cell-surface or organelle-specific proteins, and post-translational modifications such as phosphorylation or glycosylation.

Human Mesenchymal Stem Cell SILAC Kit
Kit and specialized media for stable isotope labeling using amino acids in cell culture (SILAC) involving human mesenchymal stem cells. 

Mouse Embryonic Stem Cell SILAC Kit
Kit and specialized media for stable isotope labeling using amino acids in cell culture (SILAC) involving mouse embryonic stem cells. 

References

1. Amanchy, R. et al. (2005). Science STKE 267: 1-20
2. Blagoev, B., et al. (2004). Nat Biotechnol 22(9): 1139-1145.
3. Everley, P. et al. as presented at The American Society for Biocehmistry and Molecular Biology. MCP Papers in Press. Published on July 11, 2007 as Manuscript M700057-MCP200.
4. Kratchmarova, I., et al. (2005). Science 308(5727): 1472-1477.
5. Mann, M. (2006). Functional and Quantitative Proteomics using SILAC.  Nat Rev Mol Cell Biol 7(12): 952-958.
6. Ong, S. E., Blagoev, B., Kratchmarova, I., Kristensen, D.B., Steen, H., Pandey, A., and Mann, M. (2002). Stable Isotope Labeling by Amino Acids in Cell Culture, SILAC, as a Simple and Accurate Approach to Expression Proteomics. Mol Cell Proteomics 1(5): 376-386.
7. Selbach, M. and Mann, M. (2006). Nat. Methods 3(12): 981-3