Wong Alex Y H, Xie Sheng, Tang Ben Zhong, Chen Sijie
Ming Wai Lau Centre for Reparative Medicine, Karolinska Institutet.
Department of Chemistry, Hong Kong Branch of Chinese National Engineering Research Center for Tissue Restoration and Reconstruction, Institute of Molecular Functional Materials, State Key Laboratory of Neuroscience, Division of Biomedical Engineering, and Division of Life Science, Hong Kong University of Science and Technology.
J Vis Exp. 2019 Apr 21(146). doi: 10.3791/58669.
Silver staining is a colorimetric technique widely used to visualize protein bands in polyacrylamide gels following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The classic silver stains have certain drawbacks, such as high background staining, poor protein recovery, low reproducibility, a narrow linear dynamic range for quantification, and limited compatibility with mass spectrometry (MS). Now, with the use of a fluorogenic Ag probe, TPE-4TA, we developed a fluorescent silver staining method for the total protein visualization in polyacrylamide gels. This new stain avoids the troublesome silver reduction step in traditional silver stains. Moreover, the fluorescent silver stain demonstrates good reproducibility, sensitivity, and linear quantification in protein detection, making it a useful and practical protein gel stain.
银染是一种比色技术，广泛用于在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）后可视化聚丙烯酰胺凝胶中的蛋白条带。经典的银染有一些缺点，如背景染色高、蛋白质回收率低、重现性差、定量的线性动态范围窄以及与质谱（MS）的兼容性有限。现在，通过使用荧光银探针TPE-4TA，我们开发了一种用于聚丙烯酰胺凝胶中总蛋白可视化的荧光银染方法。这种新的染色方法避免了传统银染中麻烦的银还原步骤。此外，荧光银染在蛋白质检测中表现出良好的重现性、灵敏度和线性定量，使其成为一种有用且实用的蛋白质凝胶染色方法。
