Shevchenko Andrej, Tomas Henrik, Havlis Jan, Olsen Jesper V, Mann Matthias
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany.
Nat Protoc. 2006;1(6):2856-60. doi: 10.1038/nprot.2006.468.
In-gel digestion of proteins isolated by gel electrophoresis is a cornerstone of mass spectrometry (MS)-driven proteomics. The 10-year-old recipe by Shevchenko et al. has been optimized to increase the speed and sensitivity of analysis. The protocol is for the in-gel digestion of both silver and Coomassie-stained protein spots or bands and can be followed by MALDI-MS or LC-MS/MS analysis to identify proteins at sensitivities better than a few femtomoles of protein starting material.
通过凝胶电泳分离的蛋白质的胶内消化是质谱驱动蛋白质组学的基石。舍甫琴科等人10年前的方法已得到优化，以提高分析速度和灵敏度。该方案适用于银染和考马斯亮蓝染色的蛋白质斑点或条带的胶内消化，后续可进行基质辅助激光解吸电离质谱（MALDI-MS）或液相色谱-串联质谱（LC-MS/MS）分析，以鉴定起始蛋白质材料低至飞摩尔级别的灵敏度的蛋白质。
