Effects of increased voltage on resolution in preparative isoelectric focusing of myoglobin varia

Electrophoresis. 2006 Sep;27(17):3325-31. doi: 10.1002/elps.200500939.

Abstract

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.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Chromatography, High Pressure Liquid
  • Electrophoresis, Polyacrylamide Gel / methods
  • Genetic Variation
  • Hydrogen-Ion Concentration
  • Isoelectric Focusing / methods*
  • Myoglobin / genetics
  • Myoglobin / isolation & purification*

Substances

  • Myoglobin