Promoted negative staining of proteins in SDS-PAGE using Eosin B compounded with magnesium chloride.
Electrophoresis
; 31(23-24): 3808-15, 2010 Dec.
Article
em En
| MEDLINE
| ID: mdl-21080370
ABSTRACT
In this study, we describe an effective visualizing technique for proteins in SDS-PAGE based on the organic dye, Eosin B, the sensitivity of which can be further strengthened by the addition of magnesium to the staining solution after electrophoresis. The newly developed protocol is low in cost and easily performed compared with the common methods for protein analysis in 1-D and 2-D gels. It provides a much better sensitivity (0.2 ng of single protein band) than that of imidazole-zinc negative stain for fixing and staining within 1 h, and an excellent performance in terms of compatibility with MALDI-TOF MS. The results show that similar identification scores and numbers of matched peptides were obtained by both methods. Furthermore, the effects of different metal salts on the quality of protein visualization by Eosin B were also investigated. Because of its sensitivity, stability, and safety, this stain may be a more practical method for protein determination in the routine laboratory.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas
/
Cloreto de Magnésio
/
Coloração Negativa
/
Proteômica
/
Eletroforese em Gel de Poliacrilamida
/
Fluoresceínas
Tipo de estudo:
Guideline
/
Prognostic_studies
Idioma:
En
Revista:
Electrophoresis
Ano de publicação:
2010
Tipo de documento:
Article
País de afiliação:
Coréia do Sul