Prestaining of glycoproteins in SDS-PAGE via 4H-[1]-Benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide with weak influence on protein mobility

Electrophoresis. 2014 Dec;35(24):3512-7. doi: 10.1002/elps.201400351. Epub 2014 Oct 21.

Abstract

A new fluorescent prestaining method for gel-separated glycoproteins in 1D and 2D SDS-PAGE was developed by using 4H-[1]-Benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide (BH). The prestained gels were readily imaged after electrophoresis without any time-consuming steps needed for poststain. As low as 4-8 ng glycoproteins (transferrin, α1-acid glycoprotein) could be selectively detected, which is comparable to the most commonly used Pro-Q Emerald 488 glycoprotein stain. In addition, subsequent study of deglycosylation, glycoprotein affinity chromatography, and LC-MS/MS analysis were performed to confirm the specificity of the newly developed method. As a result, BH prestain provides a new choice for quick, sensitive, specific, economical, and MS compatible visualization of gel-separated glycoproteins.

Keywords: 4H-[1]-Benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide; Glycoprotein detection; Prestain.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Blood Proteins / analysis
  • Blood Proteins / chemistry*
  • Coloring Agents / chemistry*
  • Electrophoresis, Gel, Two-Dimensional
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Glycoproteins / analysis
  • Glycoproteins / chemistry*
  • Humans
  • Molecular Sequence Data
  • Thiophenes / chemistry*

Substances

  • Blood Proteins
  • Coloring Agents
  • Glycoproteins
  • Thiophenes
  • thiophene-2-carboxylic acid hydrazide