Mass spectrometric identification of proteins from silver-stained polyacrylamide gel: a method for the removal of silver ions to enhance sensitivity

Electrophoresis. 1999 Mar;20(3):601-5. doi: 10.1002/(SICI)1522-2683(19990301)20:3<601::AID-ELPS601>3.0.CO;2-6.

Abstract

Mass spectrometry is a powerful technique for the identification of proteins at nanogram quantities. However, some degree of sample preparation prior to mass spectrometry is required, and silver-stained protein gel samples are most problematic. Here we report our strategy to obtain peptide mass profiles from silver-stained protein gel samples from one- or two-dimensional gels by destaining prior to enzymatic digestion. This study demonstrates that by using the destaining method, the sensitivity and quality of mass spectra is increased for matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometric analysis, permitting more proteins to be identified by peptide mass database analysis.

MeSH terms

  • Acrylic Resins*
  • Electrophoresis, Gel, Two-Dimensional / methods
  • Electrophoresis, Polyacrylamide Gel / methods
  • Ions
  • Proteins / analysis*
  • Sensitivity and Specificity
  • Silver Staining / methods*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*

Substances

  • Acrylic Resins
  • Ions
  • Proteins
  • polyacrylamide gels