Pozniak Yair, Geiger Tamar
The Department of Human Molecular Genetics and Biochemistry, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
Methods Mol Biol. 2014;1188:281-91. doi: 10.1007/978-1-4939-1142-4_20.
Stable isotope labeling with amino acids in cell culture (SILAC) is considered the most accurate method for proteome quantification by mass spectrometry. As it relies on active protein translation, it was traditionally limited to cells in culture and was not applicable to tissues. We have previously developed the super-SILAC mix, which is a mixture of several cell lines that serves as an internal spike-in standard for the study of human tumor tissue. The super-SILAC mix greatly improves the quantification accuracy while lowering error rates, and it is a simple, economic, and robust technique. Here we describe the design and application of super-SILAC to a broad range of biological systems, for basic biological research as well as clinical one.
细胞培养中氨基酸稳定同位素标记（SILAC）被认为是通过质谱进行蛋白质组定量的最准确方法。由于它依赖于活跃的蛋白质翻译，传统上仅限于培养中的细胞，不适用于组织。我们之前开发了超级SILAC混合物，它是几种细胞系的混合物，用作研究人类肿瘤组织的内部插入标准。超级SILAC混合物在降低错误率的同时大大提高了定量准确性，并且是一种简单、经济且可靠的技术。在此我们描述超级SILAC在广泛生物系统中的设计和应用，用于基础生物学研究以及临床研究。
