Quantitation of mucus glycoproteins blotted onto nitrocellulose membranes

Anal Biochem. 1989 Oct;182(1):160-4. doi: 10.1016/0003-2697(89)90735-5.

Abstract

A sensitive assay for mucus glycoproteins (mucins) and fragments thereof is presented. The macromolecules are blotted onto nitrocellulose membranes and visualized using a periodate-Schiff (PAS) reaction and the color yield quantitated with an image analysis system used as a reflectance densitometer. At least 50 ng of the macromolecules was detected. "Whole" mucins and subunits were assayed on 0.2-micron pore size nitrocellulose membranes whereas immobilization of the high-molecular-weight mucin glycopeptides (Mr 300-500,000) required pretreatment of membranes with poly-L-lysine. Binding of the glycopeptides to the polylysine-treated membranes was found to decrease with increasing salt concentration suggesting an electrostatic interaction. The data obtained with this method and a solution PAS assay are in good agreement but the former is more sensitive and can be performed on samples dissolved in chaotropic solvents.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Collodion
  • Colorimetry
  • Electrophoresis / methods
  • Membranes, Artificial
  • Microchemistry / methods
  • Mucins / analysis*
  • Peptide Fragments / analysis
  • Periodic Acid-Schiff Reaction
  • Reference Standards

Substances

  • Membranes, Artificial
  • Mucins
  • Peptide Fragments
  • Collodion