Kurien Biji T, Scofield R Hal
Department of Veterans Affairs, University of Oklahoma Health Sciences Center, Oklahoma Medical Research Foundation, Oklahoma City, OK, USA.
Methods Mol Biol. 2012;869:471-9. doi: 10.1007/978-1-61779-821-4_41.
Coomassie Brilliant Blue is commonly used for the detection of proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, owing to its reliability and simplicity. Here, we report dramatically decreased protein staining and destaining time, as well as significantly increased detection sensitivity with the application of enhanced heat. The staining time was 5 min at 55, 62.5, or 70°C for a 1.5-mm gel, while it took 45, 45, and 20 min, respectively, for destaining. The staining time could be reduced to 1 min for a 0.8 mm gel stained at 65°C, to 2 min at 60°C and 5 min at 55°C. The destaining of proteins analyzed on a 0.8 mm gel could be accomplished in 8, 15, and 20 min at 65, 60, and 55°C, respectively. Application of heat, thus, enables proteins to be stained and destained rapidly, as well as enhancing detection sensitivity.
考马斯亮蓝由于其可靠性和简便性，常用于十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳中蛋白质的检测。在此，我们报告了通过施加增强热量，蛋白质染色和脱色时间大幅减少，以及检测灵敏度显著提高。对于1.5毫米厚的凝胶，在55、62.5或70°C下染色时间为5分钟，而脱色分别需要45、45和20分钟。对于0.8毫米厚的凝胶，在65°C下染色时间可减少至1分钟，在60°C下为2分钟，在55°C下为5分钟。在0.8毫米厚的凝胶上分析的蛋白质脱色，在65、60和55°C下分别可在8、15和20分钟内完成。因此，加热能够使蛋白质快速染色和脱色，同时提高检测灵敏度。
