Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Subscribe

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).

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Absolute Quantitation of Isoforms of Post-translationally Modified Proteins in Transgenic Organism.
Y. Li, Y. Shu, C. Peng, L. Zhu, G. Guo, and N. Li (2012)
Mol. Cell. Proteomics 11, 272-285
   Abstract »    Full Text »    PDF »
Using Extracellular Matrix Proteomics to Understand Left Ventricular Remodeling.
M. L. Lindsey, S. T. Weintraub, and R. A. Lange (2012)
Circ Cardiovasc Genet 5, o1-o7
   Full Text »    PDF »
Proteogenomic Analysis of Mycobacterium tuberculosis By High Resolution Mass Spectrometry.
D. S. Kelkar, D. Kumar, P. Kumar, L. Balakrishnan, B. Muthusamy, A. K. Yadav, P. Shrivastava, A. Marimuthu, S. Anand, H. Sundaram, et al. (2011)
Mol. Cell. Proteomics 10, M111.011627
   Abstract »    Full Text »    PDF »
Global Phosphoproteomics Reveals Crosstalk Between Bcr-Abl and Negative Feedback Mechanisms Controlling Src Signaling.
L. Rubbi, B. Titz, L. Brown, E. Galvan, E. Komisopoulou, S. S. Chen, T. Low, M. Tahmasian, B. Skaggs, M. Muschen, et al. (2011)
Science Signaling 4, ra18
   Abstract »    Full Text »    PDF »
Fluorescence two-dimensional difference gel electrophoresis for biomaterial applications.
L. E. McNamara, M. J. Dalby, M. O. Riehle, and R. Burchmore (2010)
J R Soc Interface 7, S107-S118
   Abstract »    Full Text »    PDF »
Quantitative Proteomics Analysis of Macrophage Rafts Reveals Compartmentalized Activation of the Proteasome and of Proteasome-mediated ERK Activation in Response to Lipopolysaccharide.
S. Dhungana, B. A. Merrick, K. B. Tomer, and M. B. Fessler (2009)
Mol. Cell. Proteomics 8, 201-213
   Abstract »    Full Text »    PDF »
Identification of Secreted Proteins that Mediate Cell-Cell Interactions in an In vitro Model of the Lung Cancer Microenvironment.
L. Zhong, J. Roybal, R. Chaerkady, W. Zhang, K. Choi, C. A. Alvarez, H. Tran, C. J. Creighton, S. Yan, R. M. Strieter, et al. (2008)
Cancer Res. 68, 7237-7245
   Abstract »    Full Text »    PDF »
Abraxas and RAP80 Form a BRCA1 Protein Complex Required for the DNA Damage Response.
B. Wang, S. Matsuoka, B. A. Ballif, D. Zhang, A. Smogorzewska, S. P. Gygi, and S. J. Elledge (2007)
Science 316, 1194-1198
   Abstract »    Full Text »    PDF »
Biomarker Discovery from Pancreatic Cancer Secretome Using a Differential Proteomic Approach.
M. Gronborg, T. Z. Kristiansen, A. Iwahori, R. Chang, R. Reddy, N. Sato, H. Molina, O. N. Jensen, R. H. Hruban, M. G. Goggins, et al. (2006)
Mol. Cell. Proteomics 5, 157-171
   Abstract »    Full Text »    PDF »

To Advertise     Find Products


Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882