Stochaj Wayne R, Berkelman Tom, Laird Nancy
CSH Protoc. 2007 May 1;2007:pdb.prot4742. doi: 10.1101/pdb.prot4742.
INTRODUCTIONThe ammoniacal silver staining method is one of the most sensitive methods used to detect proteins on an SDS-PAGE gel. However, this and other standard silver staining methods are not compatible with mass spectrometry (MS), which is fast becoming the best way to identify proteins isolated on 2D gels. Because the proteins in gels to be analyzed by mass spectroscopy cannot be modified, many of the common sensitizing agents (e.g., glutaraldehyde and strong oxidizing agents) cannot be used. This method is compatible with MALDI and ESI-MS, and it shows an increased ability to deal with semipreparative protein loads without negative staining as compared with other silver staining methods. However, this process is less sensitive than standard silver staining methods.
引言
氨性银染色法是用于检测SDS-PAGE凝胶上蛋白质的最灵敏方法之一。然而，这种方法以及其他标准银染色方法与质谱分析（MS）不兼容，而质谱分析正迅速成为鉴定二维凝胶上分离出的蛋白质的最佳方法。由于要通过质谱分析的凝胶中的蛋白质不能被修饰，许多常见的敏化剂（如戊二醛和强氧化剂）不能使用。该方法与基质辅助激光解吸电离质谱（MALDI）和电喷雾电离质谱（ESI-MS）兼容，并且与其他银染色方法相比，它在处理半制备性蛋白质上样量时具有更强的能力，且无需负染。然而，该方法的灵敏度低于标准银染色方法。
