Mass Spectrometry-Based Proteomics in Molecular Diagnostics: Discovery of Cancer Biomarkers Using Tissue Culture
A schematic overview of SILAC. Cells are grown in normal and heavy amino acids containing media for 6 generations to achieve maximal incorporation of heavy amino acids. The proteins are extracted from both populations of cells and mixed in equal proportion and then subjected to either in-gel or in-solution digestion. Relative abundance of the digested peptides is determined from the ratio of heavy-to-light peptide signals as obtained from MS.