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Rapid Electrophoretic Staining and Destaining of Polyacrylamide Gels
Coomassie brilliant blue (CBB) dyes have been commonly used for the staining of protein bands in polyacrylamide gel electrophoresis (PAGE) gels. However, the staining and destaining of CBB dyes are time-consuming, and the use of methanol is hazardous to one’s health. I introduce a rapid electrophore...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MDPI
2018
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6526456/ https://www.ncbi.nlm.nih.gov/pubmed/31164558 http://dx.doi.org/10.3390/mps1020013 |
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author | Motojima, Fumihiro |
author_facet | Motojima, Fumihiro |
author_sort | Motojima, Fumihiro |
collection | PubMed |
description | Coomassie brilliant blue (CBB) dyes have been commonly used for the staining of protein bands in polyacrylamide gel electrophoresis (PAGE) gels. However, the staining and destaining of CBB dyes are time-consuming, and the use of methanol is hazardous to one’s health. I introduce a rapid electrophoretic destaining method using a semi-dry transfer unit and a high current power supply. In this method, ethanol was used instead of the hazardous methanol. Most of the protein bands became visible in 30 min. After a secondary destaining step, residual CBB was completely destained. The detection limit for a tested protein (5 ng) was higher than that of the conventional method. Therefore, this method is superior in its speed, safety, low cost, and sensitivity. |
format | Online Article Text |
id | pubmed-6526456 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-65264562019-05-31 Rapid Electrophoretic Staining and Destaining of Polyacrylamide Gels Motojima, Fumihiro Methods Protoc Protocol Coomassie brilliant blue (CBB) dyes have been commonly used for the staining of protein bands in polyacrylamide gel electrophoresis (PAGE) gels. However, the staining and destaining of CBB dyes are time-consuming, and the use of methanol is hazardous to one’s health. I introduce a rapid electrophoretic destaining method using a semi-dry transfer unit and a high current power supply. In this method, ethanol was used instead of the hazardous methanol. Most of the protein bands became visible in 30 min. After a secondary destaining step, residual CBB was completely destained. The detection limit for a tested protein (5 ng) was higher than that of the conventional method. Therefore, this method is superior in its speed, safety, low cost, and sensitivity. MDPI 2018-04-10 /pmc/articles/PMC6526456/ /pubmed/31164558 http://dx.doi.org/10.3390/mps1020013 Text en © 2018 by the author. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Protocol Motojima, Fumihiro Rapid Electrophoretic Staining and Destaining of Polyacrylamide Gels |
title | Rapid Electrophoretic Staining and Destaining of Polyacrylamide Gels |
title_full | Rapid Electrophoretic Staining and Destaining of Polyacrylamide Gels |
title_fullStr | Rapid Electrophoretic Staining and Destaining of Polyacrylamide Gels |
title_full_unstemmed | Rapid Electrophoretic Staining and Destaining of Polyacrylamide Gels |
title_short | Rapid Electrophoretic Staining and Destaining of Polyacrylamide Gels |
title_sort | rapid electrophoretic staining and destaining of polyacrylamide gels |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6526456/ https://www.ncbi.nlm.nih.gov/pubmed/31164558 http://dx.doi.org/10.3390/mps1020013 |
work_keys_str_mv | AT motojimafumihiro rapidelectrophoreticstaininganddestainingofpolyacrylamidegels |