Fully reversible procedure for silver staining improves densitometry of complex mixtures of biopolymers resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
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.
Rizzi
Corrado
C
Dipartimento Scientifico e Tecnologico, Università di Verona, Verona, Italy.
Rossini
Katia
K
Bruson
Adonella
A
Sandri
Marco
M
Dal Belin Peruffo
Angelo
A
Carraro
Ugo
U
D016477:Artifacts; D001704:Biopolymers / Q000032:analysis* / Q000737:chemistry; D003720:Densitometry / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008970:Molecular Weight; D012834:Silver / Q000737:chemistry; D016622:Silver Staining / Q000379:methods*
D001704:Biopolymers; D012834:Silver