Kimmel Center for Biology and Medicine at the Skirball Institute and Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, 540 First Avenue, New York, NY, 10016, USA.
Adv Exp Med Biol. 2014;806:93-106. doi: 10.1007/978-3-319-06068-2_5.
Stable isotope labeling by amino acids in cell culture (SILAC) is a powerful approach for high-throughput quantitative proteomics. SILAC allows highly accurate protein quantitation through metabolic encoding of whole cell proteomes using stable isotope labeled amino acids. Since its introduction in 2002, SILAC has become increasingly popular. In this chapter we review the methodology and application of SILAC, with an emphasis on three research areas: dynamics of posttranslational modifications, protein-protein interactions, and protein turnover.
稳定同位素标记的氨基酸在细胞培养中的应用(SILAC)是一种高通量定量蛋白质组学的强大方法。SILAC 通过使用稳定同位素标记的氨基酸对整个细胞蛋白质组进行代谢编码,从而实现高度准确的蛋白质定量。自 2002 年引入以来,SILAC 越来越受欢迎。本章综述了 SILAC 的方法学和应用,重点介绍了三个研究领域:翻译后修饰的动态、蛋白质-蛋白质相互作用和蛋白质周转。