Elimination of basic gaps at high pH values in 2-DE

Proteomics. 2009 Dec;9(24):5558-61. doi: 10.1002/pmic.200900091.

Abstract

The appearance of gaps, vertical lanes lacking protein spots at the cathodic end of 2-D maps generated with wide range IPG-strips exceeding a pH value of 9, is shown to depend on the electro-osmotic transport of water into the IPG-strip. Substitution of urea solution with water is demonstrated to increase the hydrolysis rate of polyacrylamide in IPG-strips explaining the gap formation. The use of 8 M urea or thiourea/urea solutions in the electrode wick enables overnight focusing without the appearance of gaps.

MeSH terms

  • Electrophoresis, Gel, Two-Dimensional / methods*
  • Escherichia coli / chemistry
  • Escherichia coli Proteins / analysis
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Proteins / analysis*
  • Urea / chemistry

Substances

  • Escherichia coli Proteins
  • Proteins
  • Urea