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Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics (2002)
| Venue: | Mol. Cell. Proteomics |
| Citations: | 593 - 23 self |
BibTeX
@ARTICLE{Ong02stableisotope,
author = {Shao-en Ong and Blagoy Blagoev and Irina Kratchmarova and Dan Bach Kristensen and Akhilesh P and Matthias Mann},
title = {Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics},
journal = {Mol. Cell. Proteomics},
year = {2002},
pages = {376--386}
}
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Abstract
The abbreviations used are: SILAC: Stable isotope labeling by amino acids in cell culture, 2DE: two dimensional (isoelectric focusing/SDS-PAGE) gel electrophoresis: ICATTM: isotope-coded affinity tag; MS: mass spectrometry; MALDI-TOF: matrix assisted laser desorption ionization-time of flight; PMF: peptide mass fingerprinting; LC-MS: liquid chromatography-MS; Copyright 2002 by The American Society for Biochemistry and Molecular Biology, Inc. Quantitative proteomics has traditionally been performed by 2D gel electrophoresis but recently, mass spectrometric methods based on stable isotope quantitation have shown great promise for the simultaneous and automated identification and quantitation of complex protein mixtures. Here we describe a method, termed SILAC for Stable Isotope Labeling by Amino acids in Cell culture, for the in vivo incorporation of specific amino acids into all mammalian proteins. Mammalian cell lines are grown in media lacking a standard essential amino acid but supplemented with a non-radioactive, isotopically labeled form of that amino acid, in this case deuterated leucine (Leu-d3). We find that growth of cells maintained in these media is no different from growth in normal media as evidenced by cell morphology,
Keyphrases
amino acid stable isotope labeling cell culture accurate approach gel electrophoresis vivo incorporation peptide mass fingerprinting mass spectrometry normal medium isotope-coded affinity tag quantitative proteomics mammalian cell line specific amino acid liquid chromatography-ms standard essential amino acid american society molecular biology desorption ionization-time automated identification great promise mammalian protein stable isotope quantitation mass spectrometric method cell morphology complex protein mixture labeled form
