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

[1] G Romain, L Cédric, P Jean-Max, et al. Quantitative phosphoproteomics revealed interplay between Syk and Lyn in the resistance to nilotinib in chronic myeloid leukemia cells. Blood 2011, 118(8):2211-2221
[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