Effects of increased voltage on resolution in preparative isoelectric focusing of myoglobin varia.
IEF is a powerful technique which separates proteins and other amphoteric solutes in a pH gradient according to their pI'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.
Bottenus
Danny
D
Chemical Engineering, Washington State University, Pullman, 99164-2710, USA. bottenus@wsu.edu
Leatzow
Dan
D
Ivory
Cornelius
C
D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D014644:Genetic Variation; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods*; D009211:Myoglobin / Q000235:genetics / Q000302:isolation & purification*
D009211:Myoglobin