A SILAC approach involving incorporation of tyrosine labeled with nine carbon-13 atoms (C) instead of the normal carbon-12 (C) has been utilized to study tyrosine kinase substrates in signaling pathways. SILAC has emerged as a very powerful method to study cell signaling, post translation modifications such as phosphorylation, protein–protein interaction and regulation of gene expression. In addition, SILAC has become an important method in secretomics, the global study of secreted proteins and secretory pathways. It can be used to distinguish between proteins secreted by cells in culture and serum contaminants. Standardized protocols of SILAC for various applications have also been published.