采用聚丙烯酰胺凝胶电泳前染色法,考察了多种类型活性染料与牛血清白蛋白共价染色的条件。实验发现,活性蓝KN-R型标记效果最佳,其优化标记条件为:pH10.0,60℃,50min,蛋白检出限达200ng,重复性和测试范围内的定量性较好,可在电泳时直接实时观测,作为蛋白标示剂有一定的应用前景。
Reactive dyes are introduced to label bovine serum albumin (BSA) before native polyacrylamide gel electrophoresis.The results show that blue KN-R is the best dye in protein staining and the optimum reaction conditions are pH 10.0,60 ℃,50 min.The excellent reproducibility and stoichiometry within the test scope were obtained and the detection limit to BSA was 200 ng.The method is sensitive,simple,direct and convenient to be observed by naked eyes in real-time style and has good application prospects for the use as protein label agent.