An ultrasensitive stain for negative protein detection in SDS-PAGE via 4',5'-Dibromofluorescein

J Proteomics. 2017 Aug 8:165:21-25. doi: 10.1016/j.jprot.2017.06.014. Epub 2017 Jun 19.

Abstract

A highly sensitive method for brief and economical staining of proteins in 1-D and 2-D sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) by 4',5'-Dibromofluorescein (DBF) was developed in this study. Down to 0.025-0.05ng protein could be detected within 10min (only 2 steps) by DBF stain, which is approximately 10-fold more sensitive than those of Eosin Y (EY) and SYPRO Ruby stains, and 20-fold more sensitive than that of imidazole-zinc (IZ) negative stain. In addition, the LC-MS/MS results indicated that the newly developed staining method is compatible with the downstream protein identification. Moreover, the mechanism of DBF stain was investigated by molecular docking.

Keywords: 4′,5′-Dibromofluorescein; Negative stain; Protein detection; Proteomics; SDS-PAGE.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Electrophoresis, Polyacrylamide Gel / methods*
  • Fluoresceins / chemistry
  • Fluoresceins / standards*
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / standards*
  • Molecular Docking Simulation
  • Proteins / analysis*
  • Staining and Labeling / standards*
  • Time Factors

Substances

  • 4,5-dibromofluorescein
  • Fluoresceins
  • Fluorescent Dyes
  • Proteins