Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autor principal: Motojima, Fumihiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
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
_version_ 1783419898197704704
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