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A Modified CBBR-250 Microwave-Assisted Staining and Destaining Method on SDS-PAGE Gels

机译:一种改进的CBBR-250微波辅助SDS-PAGE凝胶染色和脱色方法

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The application of microwave-assisted technologies has drastically reduced overall staining and destaining times in CBB R-250 staining on SDS-polyacrylamide gels but accurate temperature control and reproducibility for conventional commercial microwaves are not satisfactory. The objective of this study is to improve the existing procedures and develop an optimal method for using microwave-assisted technologies. Researchers adjusted the component of staining and destaining solutions and attempted to control the microwave temperature by shortening time, the staining and destaining could be completed within 30 min and resulted in dark purple-blue protein bands. In addition, the SDS-polyacrylamide gel displayed a transparent gel background. The detection limit for bovine serum albumin and fetuin-A on the SDS-polyacrylamide gels was 0.01 μg. These results showed that the modified method required less time. Furthermore, the modified procedure minimized the use of harmful organic solvents due to ethanol instead of methanol, methanol is more toxic than ethanol yet it maintained a high sensitivity for protein staining on SDS-polyacrylamide gels.
机译:微波辅助技术的应用已大大减少了SDS-聚丙烯酰胺凝胶上CBB R-250染色的总体染色和脱色时间,但是常规商业微波的精确温度控制和可重复性并不令人满意。这项研究的目的是改善现有程序,并开发一种使用微波辅助技术的最佳方法。研究人员调整了染色和脱色溶液的成分,并试图通过缩短时间来控制微波温度,染色和脱色可以在30分钟内完成,并产生深紫色蓝色蛋白带。另外,SDS-聚丙烯酰胺凝胶显示出透明的凝胶背景。 SDS-聚丙烯酰胺凝胶上牛血清白蛋白和胎球蛋白-A的检出限为0.01μg。这些结果表明,改进的方法需要更少的时间。此外,由于乙醇代替了甲醇,因此经过改进的程序最大限度地减少了有害有机溶剂的使用,甲醇比乙醇更具毒性,但仍对SDS-聚丙烯酰胺凝胶上的蛋白质染色保持了高敏感性。

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