使用SILAC定量比较蛋白质组

Q1 Biochemistry, Genetics and Molecular Biology

Current Protocols in Protein Science Pub Date : 2018-09-20 DOI:10.1002/cpps.74

Jingjing Deng, Hediye Erdjument-Bromage, Thomas A. Neubert

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引用次数: 25

摘要

细胞培养中氨基酸稳定同位素标记(SILAC)作为一种定量蛋白质组学方法，自2002年由Matthias Mann的团队首次引入以来，已成为非常受欢迎的方法。这是一种代谢标记策略，在翻译过程中，同位素标记的氨基酸在体内代谢结合到蛋白质中。在自然(轻)或重氨基酸掺入后，在细胞裂解后和任何进一步处理之前立即混合差异标记的样品，这将最大限度地减少平行处理不同样品造成的定量误差。在本单元中，我们描述了用于非分裂细胞(如神经元)的基本双工SILAC和三工SILAC的协议，以及用于测量新合成蛋白质的量。©2018 by John Wiley &儿子,Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Quantitative Comparison of Proteomes Using SILAC

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Quantitative Comparison of Proteomes Using SILAC

Stable isotope labeling by amino acids in cell culture (SILAC) has become very popular as a quantitative proteomic method since it was firstly introduced by Matthias Mann's group in 2002. It is a metabolic labeling strategy in which isotope-labeled amino acids are metabolically incorporated in vivo into proteins during translation. After natural (light) or heavy amino acid incorporation, differentially labeled samples are mixed immediately after cell lysis and before any further processing, which minimizes quantitative errors caused by handling different samples in parallel. In this unit, we describe protocols for basic duplex SILAC, triplex SILAC for use in nondividing cells such as neurons, and for measuring amounts of newly synthesized proteins. © 2018 by John Wiley & Sons, Inc.

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来源期刊

Current Protocols in Protein Science Biochemistry, Genetics and Molecular Biology-Biochemistry

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期刊介绍： With the mapping of the human genome, more and more researchers are exploring protein structures and functions in living organisms. Current Protocols in Protein Science provides protein scientists, biochemists, molecular biologists, geneticists, and others with the first comprehensive suite of protocols for this growing field.