Stable isotope labeling by amino acids in cell culture (SILAC)

Abstract

Quantitative proteomics has become a pivotal tool that has been applied to the investigation of many different biological processes such diverse as the detection of biomarkers in tissue samples, the regulation of cell signaling, and the characterization of protein interactions. Stable isotope labeling techniques have facilitated the precise quantitation of changes in protein abundance by mass spectrometry. Among different choices, Stable Isotope Labeling by Amino acids in Cell culture (SILAC) is an easy and reliable method for unbiased comparative proteomic experiments, which has been employed to study post-translational modifications such as protein phosphorylation and methylation, to characterize signaling pathways and to determine specific protein interactions. Here we describe detailed procedures for SILAC experiments in mammalian and yeast cells.