Real-time monitoring of polyacrylamide gel electrophoresis by the shadowgraph technique. (10th November 2022)

• Record Type: Journal Article
• Authors: Murakami, Satoshi; Adachi, Hiroaki; Matsumura, Hiroyoshi; Takano, Kazufumi; Inoue, Tsuyoshi; Mori, Yusuke
• Is Part Of: Journal of biochemistry. Volume 173:Number 2(2023)
• Page Start: 65
• Page End: 72
• Publication Date: 2022-11-10
• Subjects: shadowgraph -- SDS-PAGE -- protein analysis -- polyacrylamide gel electrophoresis -- direct observation -- Abbreviations: CBB, Coomassie Brilliant Blue; PAGE, Polyacrylamide gel electrophoresis; SDS, Sodium dodecyl sulphate; Biochemistry -- Periodicals; Biochemistry -- Periodicals; Electronic journals; 572.05
• DOI: 10.1093/jb/mvac087
• Languages: English
• ISSNs: 0021-924X

Abstract

Abstract: 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. Graphical Abstract: