Real-time monitoring of polyacrylamide gel electrophoresis by the shadowgraph technique
Real-time monitoring of polyacrylamide gel electrophoresis by the shadowgraph technique
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阴影技术实时监测聚丙烯酰胺凝胶电泳
DOI：
10.1093/jb/mvac087
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Mori Yusuke
中科院分区：
文献类型：
--
作者：
Murakami Satoshi;Adachi Hiroaki;Matsumura Hiroyoshi;Takano Kazufumi;Inoue Tsuyoshi;Mori Yusuke
采用十二烷基硫酸钠 (SDS) 和考马斯亮蓝 (CBB) 染色的聚丙烯酰胺凝胶电泳 (PAGE) 广泛用于蛋白质研究，并且需要时间进行电泳、染色和脱色。由于凝胶中电泳的蛋白条带在大多数情况下是不可见的，因此只有整个过程完成后才能观察到结果。在这项研究中，阴影图用于在电泳过程中检测蛋白质等生物分子。简单的光学系统和摄像头可实时监测聚丙烯酰胺凝胶中单个蛋白质条带的迁移和分离，无需染色。能见度足够高，可以看到蛋白质以外的物质，例如 DNA。该方法在电泳的早期阶段立即提供蛋白质谱。取消染色和脱色步骤将有助于节省研究人员的时间。该方法也是环保的，将有助于减少含有合成染料的废液的产生。
Polyacrylamide gel electrophoresis (PAGE) with sodium dodecyl sulphate (SDS) and Coomassie brilliant blue (CBB) staining is widely used in protein research and requires time for electrophoresis, staining and destaining. Because the protein bands electrophoresed in the gel are invisible in most cases, the results cannot be observed until the whole process is complete. In this study, shadowgraph was applied to detect biomolecules such as proteins during electrophoresis. A simple optical system and camera-enabled real-time monitoring of migration and separation of individual protein bands in polyacrylamide gels without staining. The visibility was high enough that it was possible to visualize substances other than proteins, such as DNA. This method provides protein profiles instantly in the early stage of electrophoresis. The elimination of the staining and destaining steps will help save researchers’ time. The method is also environmentally friendly and will help reduce the generation of waste solutions containing synthetic dyes.