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SILAC

SILAC(Stable isotope labeling with amino acids in cell culture)也叫AACT(amino acid-coded mass tagging)属于体内标记技术

技术原理

SILAC方法通过“重型”13C-或15N-标记的氨基酸在体内代谢过程中掺入到蛋白质中,等量混合各类型同位素氨基酸标记的蛋白质样本,酶解后进行质谱分析,通过比较一级质谱图中各类型同位素标记质谱峰型的面积大小进行相对定量,同时利用二级谱图鉴定蛋白质。

技术流程
技术优势

1.标记效率几乎100%,比体外标记技术标记效率高;
2.避免了样本处理过程中引入的误差;
3.所需样本明显少于体外标记。

应用领域

▶基础医学、临床诊断疾病机理机制、个性化治疗等;

▶ 生物医药:药物作用机理,药效评价,药物开发等

Chen, X., Smith, L.M., and Bradbury, E.M. (2000). Site-Specific Mass Tagging with Stable Isotopes in Proteins for Accurate and Efficient Protein Identification. Analytical Chemistry 72, 1134-1143.
Ong, S.-E., Blagoev, B., Kratchmarova, I., Kristensen, D.B., Steen, H., Pandey, A., and Mann, M. (2002). Stable Isotope Labeling by Amino Acids in Cell Culture, SILAC, as a Simple and Accurate Approach to Expression Proteomics. Molecular & Cellular Proteomics 1, 376-386.
Pan, S., Gu, S., Bradbury, E.M., and Chen, X. (2003). Single Peptide-Based Protein Identification in Human Proteome through MALDI-TOF MS Coupled with Amino Acids Coded Mass Tagging. Analytical Chemistry 75, 1316-1324.

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