SILAC

'The principle of SILAC'. Cells differentially labeled by growing them in light medium with normal arginine (Arg-0, grey colour) or medium with heavy arginine (Arg-6, red colour). Metabolic incorporation of the amino acids into the proteins results in a mass shift of the corresponding peptides. This mass shift can be detected by a mass spectrometer as indicated by the depicted mass spectra. When both samples are combined, the ratio of peak intensities in the mass spectrum reflects the relative protein abundance. In this example, the labeled protein has the same abundance in both samples (ratio 1).

'SILAC (Stable isotope labelling with amino acids in cell culture)' is a mass spectrometry-based technique developed in the group of Matthias Mann to detect differences in protein abundance between two samples (Ong et al., 2002). Two populations of cells are cultivated in cell culture. One of the cell populations is fed with growth medium containing normal amino acids. In contrast, the growth medium of the second cell population contains amino acids labeled with stable (non-radioactive) heavy isotopes. For example, the medium can contain arginine labeled with six carbon-13 atoms (13C) instead of the normal carbon-12 (12C). When the cells are growing in this medium, they incorporate the heavy arginine into all of their proteins. Therefore, all of the arginine containing peptides are now 6 Da heavier than their normal counterparts. The trick is that the proteins from both cell populations can be combined and analyzed together by mass spectrometry. Pairs of chemically identical peptides of different stable-isotope composition can be differentiated in a mass spectrometer owing to their mass difference. The ratio of peak intensities in the mass spectrum for such peptide pairs accurately reflects the abundance ratio for the two proteins. SILAC has emerged as a very powerful method to study cell signaling and protein-protein interaction.

Contents
References
See also
External links

References



★ Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics, Ong SE, Blagoev B, Kratchmarova I, Kristensen DB, Steen H, Pandey A, Mann M, , , Molecular & Cell Proteomics,

★ Properties of 13C-substituted arginine in stable isotope labeling by amino acids in cell culture (SILAC), Ong SE, Kratchmarova I, Mann M, , , Journal of Proteome Research,

★ Mass spectrometric-based approaches in quantitative proteomics, Ong SE, Foster LJ, Mann M, , , Methods,

★ A practical recipe for stable isotope labeling by amino acids in cell culture (SILAC), Ong SE, Mann M, , , Nature Protocols,

★ Quantitative proteomics to study mitogen-activated protein kinases, Blagoev B, Mann M, , , Methods,

★ Stable isotope labeling by amino acids in cell culture for quantitative proteomics, Ong SE, Mann M, , , Methods in Molecular Biology,

See also



★ Cell signaling

★ mass spectrometry

★ Protein-protein interaction

★ Proteomics

External links



★ Mann Lab

★ Pandey Lab

★ CEBI

★ A web resource for SILAC

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