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.
| Original language | English |
|---|---|
| Pages (from-to) | 160-4 |
| Number of pages | 5 |
| Journal | Analytical Biochemistry |
| Volume | 182 |
| Issue number | 1 |
| Publication status | Published - Oct 1989 |
Keywords
- Collodion
- Colorimetry
- Electrophoresis/methods
- Membranes, Artificial
- Microchemistry/methods
- Mucins/analysis
- Peptide Fragments/analysis
- Periodic Acid-Schiff Reaction
- Reference Standards