Fluorescent Silver Staining of Proteins in Polyacrylamide Gels

J Vis Exp. 2019 Apr 21:(146). doi: 10.3791/58669.

Abstract

Silver staining is a colorimetric technique widely used to visualize protein bands in polyacrylamide gels following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The classic silver stains have certain drawbacks, such as high background staining, poor protein recovery, low reproducibility, a narrow linear dynamic range for quantification, and limited compatibility with mass spectrometry (MS). Now, with the use of a fluorogenic Ag+ probe, TPE-4TA, we developed a fluorescent silver staining method for the total protein visualization in polyacrylamide gels. This new stain avoids the troublesome silver reduction step in traditional silver stains. Moreover, the fluorescent silver stain demonstrates good reproducibility, sensitivity, and linear quantification in protein detection, making it a useful and practical protein gel stain.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Acrylic Resins*
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Fluorescent Dyes
  • Proteins / analysis*
  • Proteins / chemistry
  • Reproducibility of Results
  • Silver Staining / methods*

Substances

  • Acrylic Resins
  • Fluorescent Dyes
  • Proteins
  • polyacrylamide gels