Institute of Hematology and Blood Transfusion, Prague, Czech Republic.
Electrophoresis. 2013 Jul;34(13):1972-5. doi: 10.1002/elps.201300087.
Several new fast staining protocols for the visualization of proteins separated by SDS-PAGE utilizing Coomassie Blue staining (CBS) have been described in literature. The sensitivity of a newly designed staining protocol is usually estimated using 1D SDS-PAGE of serially diluted protein samples. However, this approach is not predictive and satisfactory for 2D SDS-PAGE capable of resolving hundreds or thousands of different proteins in a single analysis. In this work, a new fast staining protocol recently introduced by Dong et al. (PLoS One 2011, 6, e22394) was compared to colloidal CBS. The number of detectable spots in 2D SDS-PAGE of identical blood plasma samples in repeated runs was chosen as a sensitivity criterion. Further, the influence of gel boiling on the subsequent protein identification by MS was investigated. In spite of its advantages, the staining protocol according to Dong et al. (PLoS One 2011, 6, e22394) seems to be less sensitive than colloidal Coomassie staining when the number of detected spots is the evaluating criterion. No obvious influence of gel boiling on the protein identification was observed.
已有文献中描述了几种利用考马斯亮蓝染色（CBS）对 SDS-PAGE 分离的蛋白质进行快速染色的新方案。新设计的染色方案的灵敏度通常使用连续稀释的蛋白质样品的 1D SDS-PAGE 来估计。然而，这种方法对于能够在单次分析中解析数百或数千种不同蛋白质的 2D SDS-PAGE 来说并不具有预测性和令人满意。在这项工作中，Dong 等人最近引入的一种新的快速染色方案（PLoS One 2011, 6, e22394）与胶体 CBS 进行了比较。选择在重复运行中相同的血浆样品的 2D SDS-PAGE 中可检测到的斑点数量作为灵敏度标准。此外，还研究了凝胶煮沸对随后通过 MS 进行蛋白质鉴定的影响。尽管具有优势，但当检测到的斑点数量作为评估标准时，根据 Dong 等人的方案（PLoS One 2011, 6, e22394）似乎不如胶体考马斯亮蓝染色灵敏。未观察到凝胶煮沸对蛋白质鉴定的明显影响。
