技术原理
研究细胞模型中HAT、HDAC的底物或HDACi的作用靶点时,通常采用稳定同位素标记培养基中的氨基酸(Lysine),同时结合抗体富集、高分辨率质谱的方法分析不同处理细胞系中的乙酰化组,并通过肽段一级谱图获得相对定量信息。
技术优势
1. 采用体内标记技术,几乎不影响细胞的功能
2. 灵敏度高,且定量信息准确
实验流程
参考文献
[1] P Beli, N Lukashchuk, SA Wagner, et al. Proteomic Investigations Reveal a Role for RNA Processing Factor THRAP3 in the DNA Damage Response. Mol Cell 2012, 46(2):212–225
[2] Z Shimin, X Wei, J Wenqing, et al. Regulation of Cellular Metabolism by Protein Lysine Acetylation. Science 2010, 327(5968):1000-1004
[3] D Girija, L Dmitry, K Lora, et al. The pan-HDAC inhibitor vorinostat potentiates the activity of the proteasome inhibitor carfilzomib in human DLBCL cells in vitro and in vivo. Blood 2010, 115(22):4478-4487