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Effects of increased voltage on resolution in preparative isoelectric focusing of myoglobin varia
Author(s) -
Bottenus Danny,
Leatzow Dan,
Ivory Cornelius
Publication year - 2006
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/elps.200500939
Subject(s) - isoelectric focusing , myoglobin , chemistry , resolution (logic) , electrophoresis , electric field , chromatography , analytical chemistry (journal) , voltage , physics , biochemistry , enzyme , computer science , quantum mechanics , artificial intelligence
IEF is a powerful technique which separates proteins and other amphoteric solutes in a pH gradient according to their p I 's. The current work evaluates the effect on resolution of increasing electric fields in a novel preparative, vortex‐stabilized electrophoresis device. In shallow gradients spanning one pH unit, the variants of myoglobin were separated at applied voltages from 10 to 15 kV. Digital imaging of these separations indicated a 20% reduction in bandwidth and a 60% increase in resolution as the electric field strength is varied across this range. These results were confirmed by IEF‐PAGE and ion‐exchange chromatography.