title: Characterization of biotin-labeled proteoglycans by electrophoretic separation on minigels and blotting onto nylon membranes prior and after enzymatic digestion.
abstract: Biotinylated proteoglycans were separated by sodium dodecyl sulfate electrophoresis prior and after enzymatic digestion by glycan-specific enzymes using polyacrylamide minigels. The biotin-labeled compounds were blotted onto nylon membranes either by electrophoresis or by diffusion and detected by avidin-enzyme conjugates. The method allows the nonisotopic detection of native proteoglycans and core proteins. Proteoglycans can be visualized at protein amounts as low as 0.7 ng per lane. In comparison with sensitive protein stains, compounds of enzyme preparations do not interfere with bands corresponding to core proteins. Electrophoresis, blotting, and staining of up to 12 samples per gel are accomplished in less than 3 h.
authors.affiliation: Department of Clinical Chemistry and Pathobiochemistry, Medical Faculty, University of Technology, Aachen, Federal Republic of Germany.
authors.forename: G
authors.initials: G
authors.lastname: Stöcker
authors.forename: J
authors.initials: J
authors.lastname: Lückge
authors.forename: H
authors.initials: H
authors.lastname: Greiling
authors.forename: C
authors.initials: C
authors.lastname: Wagener
mesh_terms: D000596:Amino Acids / Q000032:analysis; D001011:Aorta / Q000032:analysis; D001710:Biotin; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D006595:Hexosamines / Q000032:analysis; D006801:Humans; D005779:Immunodiffusion; D008567:Membranes, Artificial; D011509:Proteoglycans / Q000032:analysis*
chemical_list: D000596:Amino Acids; D006595:Hexosamines; D008567:Membranes, Artificial; D011509:Proteoglycans; D001710:Biotin