Previsible silver staining of protein in electrophoresis gels with mass spectrometry compatibility

Anal Biochem. 2008 Dec 15;383(2):137-43. doi: 10.1016/j.ab.2008.04.048. Epub 2008 Aug 27.

Abstract

A convenient silver staining method for protein in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels is described. The method is previsible, sensitive, and mass spectrometry (MS) compatible. Two visible counter ion dyes, ethyl violet (EV) and zincon (ZC), were used in the first staining solution with a detection limit of 2 to 8 ng/band in approximately 1h. The dye-stained gel can be further stained by silver staining, which is based on acidic silver staining employing ZC with sodium thiosulfate as silver ion sensitizers. Especially, ZC has silver ion reducing power by cleavage of the diazo bond of the dye during silver reduction. The second silver staining can be completed in approximately 1h with a detection limit of 0.2 ng/band.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analytic Sample Preparation Methods / economics
  • Analytic Sample Preparation Methods / methods*
  • Animals
  • Cattle
  • Coloring Agents / chemistry
  • Costs and Cost Analysis
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Linear Models
  • Mass Spectrometry
  • Proteins / analysis*
  • Proteins / chemistry
  • Proteomics
  • Rosaniline Dyes / chemistry
  • Sensitivity and Specificity
  • Silver Staining / economics
  • Silver Staining / methods*
  • Time Factors
  • Zinc / chemistry

Substances

  • Coloring Agents
  • Proteins
  • Rosaniline Dyes
  • ethyl violet
  • Zinc