Towards stoichiometric silver staining of proteins resolved in complex two‐dimensional electrophoresis gels: Real‐time analysis of pattern development

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Abstract
Toward the ultimate goal of deriving quantitative protein data from silver-stained proteins resolved in complex two-dimensional electrophoresis gels we have performed computerized real-time analysis during pattern protein “silver stain” development. Three points emerged from this study: (i) Development time is a far more important variable than originally envisioned. (ii) Previous studies which, on the basis of a single development time, sought to relate silver stain slope and threshold properties to amino acid composition are non-informative since it will be shown that slope values change markedly with development time, (iii) The combination of real-time analysis and total protein load consideration allows, for the first time, protein “spots” to be grouped into categories, which in turn aid in quantification.
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