Real-time monitoring of polyacrylamide gel electrophoresis by the shadowgraph technique

Author:

Murakami Satoshi12ORCID,Adachi Hiroaki2ORCID,Matsumura Hiroyoshi23ORCID,Takano Kazufumi24ORCID,Inoue Tsuyoshi25ORCID,Mori Yusuke26ORCID

Affiliation:

1. Tokyo Institute of Technology Department of Life Science and Technology, , Yokohama 226-8501, Japan

2. SOSHO Inc., Suita , Osaka 565-0871, Japan

3. Ritsumeikan University Department of Biotechnology, College of Life Sciences, , Kusatsu, Shiga 525-8577, Japan

4. Graduate School of Life and Environmental Sciences , Kyoto Prefectural University, Sakyo-ku, Kyoto 606-8522, Japan

5. Graduate School of Pharmaceutical Science Division of Advance Pharmaco-Science, , Osaka University, Suita, Osaka 565-0871, Japan

6. Graduate School of Engineering , Osaka University, Suita, Osaka 565-0871, Japan

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.

Publisher

Oxford University Press (OUP)

Subject

Molecular Biology,Biochemistry,General Medicine

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3. Disc electrophoresis. I. Background and theory;Ornstein;Ann. N. Y. Acad. Sci.,1964

4. Cleavage of structural proteins during the assembly of the head of bacteriophage T4;Laemmli;Nature,1970

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