Chevallet Mireille, Luche Sylvie, Diemer Hélène, Strub Jean-Marc, Van Dorsselaer Alain, Rabilloud Thierry
CEA-Laboratoire de Biochimie et Biophysique des systèmes Intégrés, iRTSV/BBSI, CEA-Grenoble, Grenoble, France.
Proteomics. 2008 Dec;8(23-24):4853-61. doi: 10.1002/pmic.200800321.
Protein detection methods after electrophoresis have to be sensitive, homogeneous, and not to impair downstream analysis of proteins by MS. Speed, low cost, and user friendliness are also favored features. Silver staining combines many of these features, but its compatibility with MS is limited. We describe here, a new variant of silver staining that is completely formaldehyde-free. Reducing sugars in alkaline borate buffer are used as developers. While keeping the benefits of silver staining, this method is shown to afford a much better performance in terms of compatibility with MS, both in PMF by MALDI and in LC/ESI/MS/MS.
电泳后蛋白质检测方法必须具备灵敏度高、均一性好且不影响蛋白质下游质谱分析的特点。速度快、成本低和用户友好性也是其受欢迎的特性。银染法具备许多这些特性,但其与质谱的兼容性有限。我们在此描述一种完全无甲醛的新型银染法。碱性硼酸盐缓冲液中的还原糖用作显影剂。该方法在保留银染法优点的同时,在与基质辅助激光解吸电离飞行时间质谱(MALDI)的肽质量指纹图谱(PMF)以及液相色谱/电喷雾电离串联质谱(LC/ESI/MS/MS)中的质谱兼容性方面表现出更好的性能。