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Fully reversible procedure for silver staining improves densitometry of complex mixtures of biopolymers resolved by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis
Author(s) -
Rizzi Corrado,
Rossini Katia,
Bruson Adonella,
Sandri Marco,
Dal Belin Peruffo Angelo,
Carraro Ugo
Publication year - 2002
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200210)23:19<3266::aid-elps3266>3.0.co;2-l
Subject(s) - densitometry , sodium dodecyl sulfate , silver stain , polyacrylamide , chromatography , gel electrophoresis , chemistry , polyacrylamide gel electrophoresis , staining , gel electrophoresis of proteins , electrophoresis , sodium , biochemistry , microbiology and biotechnology , biology , polymer chemistry , organic chemistry , enzyme , physics , genetics , quantum mechanics
Due to its high sensitivity, silver staining is a widely popular method for the revelation of biopolymers separated by both native and denaturing electrophoresis. A step‐by‐step method for the destaining and restaining of overdeveloped/overloaded silver‐stained bands is described that is applicable to both proteins and nucleic acids. The procedure significantly improves densitometric analysis of gels that have been silver stained with either commercial kits or solutions made in‐house. The method permits reproducible densitometry of silver‐stained gels and allows quantification of both main and minor components in complex mixture of molecules resolved on the same gel slab. All steps may be interrupted and are readily reversible, allowing for facile densitometric analyses and photographic recording under optimized conditions. Furthermore, common artifacts such as differential staining of the two gel surfaces, localized uneven yellow‐ochre background, and the presence of fold marks and fingerprints can be easily removed.