Moravec Jiri, Mares Jan
Biomedical Center, Faculty of Medicine in Pilsen, Charles University, Pilsen, Czech Republic.
Electrophoresis. 2017 Dec;38(24):3100-3103. doi: 10.1002/elps.201700189. Epub 2017 Aug 30.
We introduce an optimized periodic acid-Schiff´s staining of glycoproteins on 1D electrophoretic gels. Thanks to heating in a household microwave oven the protocol of standard periodic acid-Schiff´s staining has been accelerated from 6 h to below 10 min employing standard chemistry. At the same time, we show that the microwave-assisted glycoprotein staining is at least as sensitive as the conventional approach. All glycoproteins stained by the microwave-accelerated procedure were successfully identified using MALDI TOF/TOF mass spectrometry. The ensuing reduction in gel staining time and simplification of the staining protocol should significantly increase laboratory throughput when glycoprotein detection on electrophoretic gels is required in large numbers.
我们介绍了一种在一维电泳凝胶上对糖蛋白进行优化的过碘酸-希夫氏染色方法。借助家用微波炉加热,利用标准化学方法,标准过碘酸-希夫氏染色方案从6小时加速至不到10分钟。同时,我们表明微波辅助糖蛋白染色的灵敏度至少与传统方法相当。通过微波加速程序染色的所有糖蛋白均使用基质辅助激光解吸电离飞行时间/飞行时间质谱成功鉴定。当大量需要在电泳凝胶上检测糖蛋白时,随之而来的凝胶染色时间缩短和染色方案简化应会显著提高实验室通量。