技术原理
研究细胞模型中激酶或激酶抑制剂的底物,通常采用稳定同位素标记培养基中的氨基酸(Lysine和Aginine),同时结合磷酸化富集、高分辨率质谱的方法分析不同处理细胞系中的磷酸化组,并通过肽段一级谱图获得相对定量信息。
技术优势
采用体内标记技术,几乎不影响细胞的功能
灵敏度高,且定量信息准确
实验流程
参考文献
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[2] N Thomas, P Judith Helena, CJ Tonkin, et al. Quantitative in vivo analyses reveal calcium-dependent phosphorylation sites and identifies a novel component of the Toxoplasma invasion motor complex. Plos Pathogens, 2011, 7(9):23-24
[3] JV Olsen, B Blagoev, M Mann, et al. Global, In Vivo, and Site-Specific Phosphorylation Dynamics in Signaling Networks. Cell 2006, 127(3): 635-648