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The Bio-Rad Silver Stain, Rev E


Introduction
Silver staining was first introduced as a general protein stain useful in polyacrylamide gel analysis in 1979 by Merril et al.54 This first practical PAGE silver stain was of the silver diammine type adapted from early histological silver stains. In 1981, Merril et al.55 introduced a faster, more reliable, and very sensitive silver stain derived from a photographic chemical process. Bio-Rads silver stain is based on this photochemical method.

The most important advantage of silver staining gels is the increased sensitivity obtained over other staining methods. Sensitivity is typically 50 times greater than obtained with classical Coomassie* Brilliant Blue R-250 staining. Increased sensitivity offers obvious advantages. For example, less sample is required when running gels. Also, analysis of dilute samples is possible. Protein purity can be assessed and contaminants detected more reliably. Silver stains detect a wider variety of macromolecules than does Coomassie Brilliant Blue R-250, including nucleic acids, glycoproteins, and lipoproteins.


Mechanism of Silver Staining
Mechanisms and features of silver staining have been discussed in prior reviews51, 57. The basic mechanism occurring in silver staining of macromolecules is the reduction of ionic to metallic silver. Protein bands are imaged in the gel due to differences in oxidation/reduction potentials between sites in gels occupied by protein and adjacent sites not occupied by protein. If protein-occupied sites have the higher reducing potential, then positive images are formed. Conversely, if sites unoccupied by protein have the
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