Towards stoichiometric silver staining of proteins resolved in complex two-dimensional electrophoresis gels: real-time analysis of pattern development.
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.
Rodriguez
L V
LV
University of Texas M. D. Anderson Cancer Center, Department of Molecular Pathology, Houston, TX 77030.
Gersten
D M
DM
Ramagli
L S
LS
Johnston
D A
DA
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008114:Liver Neoplasms, Experimental / Q000737:chemistry; D008297:Male; D009363:Neoplasm Proteins / Q000032:analysis*; D051381:Rats; D016622:Silver Staining
D009363:Neoplasm Proteins