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Sci. STKE, 18 January 2005
Vol. 2005, Issue 267, p. pl2
[DOI: 10.1126/stke.2672005pl2]

PROTOCOLS

Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC) for Studying Dynamics of Protein Abundance and Posttranslational Modifications

Ramars Amanchy, Dario Eluan Kalume, and Akhilesh Pandey*

McKusick-Nathans Institute of Genetic Medicine and Department of Biological Chemistry and Oncology, Johns Hopkins University, Baltimore, MD 21218, USA.

Abstract: Stable isotope labeling with amino acids in cell culture (SILAC) is a simple and straightforward approach for in vivo incorporation of a tag into proteins for relative quantitation by mass spectrometry. SILAC is a simple, yet powerful, method for investigating the dynamics of protein abundance and posttranslational modifications. Here, we provide detailed instructions for using this method to study protein complexes, protein-protein interactions, and the dynamics of protein abundance and posttranslational modifications. We expect that SILAC will become a routine technique because of its applicability to most areas of cell biology. We have also developed a Web site (http://www.silac.org) to provide researchers with updated information about this method and related resources.

*Corresponding author. Telephone, 410-502-6662; fax, 410-502-7544; e-mail, pandey{at}jhmi.edu

Citation: R. Amanchy, D. E. Kalume, A. Pandey, Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC) for Studying Dynamics of Protein Abundance and Posttranslational Modifications. Sci. STKE 2005, pl2 (2005).

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