SILAC

SILAC refers to labelling cultured cells with heavy amino acids for quantitative proteomic analysis. Labelling an entire proteome with heavy amino acids in vivo generates an ideal standard for quantitative proteomics. When a heavy labelled proteome is mixed with an unlabelled proteome then digested, every unlabelled peptide identified by the mass spectrometer can be quantified by its corresponding heavy peptide.

In SILAC, the tryptic amino acids, arginine (R) and lysine (K), contain heavy stable isotopes, so if digesting with trypsin, every peptide is labelled. This metabolic labelling strategy has been employed by hundreds of proteomic studies (see example references below). The advantage of metabolic labelling over in vitro tagging techniques is that the heavy and unlabelled samples are mixed before sample preparation, preventing variability between preparations from distorting the final quantitation results. This is especially important when extensive sample preparation (e.g. isolation of an organelle) is required.

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