Promoted negative staining of proteins in SDS-PAGE using Eosin B compounded with magnesium chloride

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...

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Bibliographic Details
Published in:Electrophoresis 2010-12, Vol.31 (23-24), p.3808-3815
Main Authors: Wang, Xu, Hwang, Sun-Young, Cong, Wei-Tao, Jin, Li-Tai, Choi, Jung-Kap
Format: Article
Language:English
Subjects:
Dyes
Electrophoresis
Electrophoresis, Polyacrylamide Gel - methods
Eosin B
Eosine I Bluish
Escherichia coli Proteins - chemistry
Fluoresceins - chemistry
Magnesium
Magnesium chloride
Magnesium Chloride - chemistry
Negative stain
Negative Staining - methods
Peptide Fragments - chemistry
PMF
Proteins
Proteins - analysis
Proteins - chemistry
Proteomics
Proteomics - methods
Stability
Staining
Visualization
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Summary: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.
ISSN:0173-0835
1522-2683
1522-2683
DOI:10.1002/elps.201000343