A novel droplet-tap sample-loading method for two-dimensional gel electrophoresis

Anal Bioanal Chem. 2006 Apr;384(7-8):1578-83. doi: 10.1007/s00216-006-0304-x. Epub 2006 Mar 23.

Abstract

A novel procedure, droplet-tap mode, has been devised for sample application for two-dimensional gel electrophoresis (2DE) expression profiles. The sample was loaded by evenly distributed tapping of droplets of the sample on to the rehydration buffer (RB) and then lowering the strip on to the solution surface. At normal loading concentrations, the number of spots obtained was increased by approximately one-third by this new approach compared with the rehydration loading procedure. The method also resulted in significantly improved resolution compared with cup loading when high concentrations of proteins were present, indicating its potential usefulness in micropreparative separation. In addition, recovery of the proteins confirmed that protein uptake was enhanced by use of this method. By enabling improved performances in 2DE, the proposed procedure has much potential for sample loading to meet the requirements of global proteome analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bombyx
  • Chemistry Techniques, Analytical / instrumentation*
  • Chemistry Techniques, Analytical / methods*
  • Electrophoresis, Gel, Two-Dimensional / instrumentation*
  • Electrophoresis, Gel, Two-Dimensional / methods*
  • Hydrogen-Ion Concentration
  • Insect Proteins / chemistry
  • Isoelectric Focusing / instrumentation
  • Isoelectric Focusing / methods
  • Protein Array Analysis / instrumentation
  • Protein Array Analysis / methods
  • Proteins / analysis*
  • Proteome
  • Proteomics / methods

Substances

  • Insect Proteins
  • Proteins
  • Proteome