Improved conditions for periodate/Schiff's base-based fluorescent staining of glycoproteins with dansylhydrazine in SDS-PAGE

Electrophoresis. 2014 May;35(10):1439-47. doi: 10.1002/elps.201300595. Epub 2014 Apr 10.

Abstract

An improved periodate/Schiff's base based fluorescent stain with dansylhydrazine (DH) for glycoproteins in 1D and 2D SDS-PAGE was described. Down to 4-8 ng of glycoproteins can be selectively detected within 2 h, which is approximately 16-fold higher than that of original protocol, but similar to that of Pro-Q Emerald 488 stain (Invitrogen, Carlsbad, USA). Furthermore, subsequent study of deglycosylation, glycoprotein affinity isolation, and LC-MS/MS analysis were performed to confirm the specificity of the improved method. As a result, improved DH stain may provide a new choice for selective, economic, MS compatible, and convenient visualization of gel-separated glycoproteins.

Keywords: Dansylhydrazine; Fluorescent stain; Glycoprotein detection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, Affinity
  • Dansyl Compounds / chemistry*
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescent Dyes / chemistry*
  • Glycoproteins / chemistry*
  • Hydrazines / chemistry*
  • Periodic Acid / chemistry*
  • Reproducibility of Results
  • Tandem Mass Spectrometry

Substances

  • Dansyl Compounds
  • Fluorescent Dyes
  • Glycoproteins
  • Hydrazines
  • Periodic Acid
  • dansyl hydrazine