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Promoted negative staining of proteins in SDS‐PAGE using Eosin B compounded with magnesium chloride
Author(s) -
Wang Xu,
Hwang SunYoung,
Cong WeiTao,
Jin LiTai,
Choi JungKap
Publication year - 2010
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201000343
Subject(s) - staining , stain , chemistry , chromatography , magnesium , silver stain , protein detection , negative stain , microbiology and biotechnology , materials science , biology , nanotechnology , organic chemistry , genetics , electron microscope , physics , optics
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.