Fluorographic detection of tritiated glycopeptides and oligosaccharides separated on polyacrylamide gels: analysis of glycans from Dictyostelium discoideum glycoproteins.
Previous workers have shown that oligosaccharides and glycopeptides can be separated by electrophoresis in buffers containing borate ions. However, normal fluorography of tritium-labeled structures cannot be performed because the glycans are soluble and can diffuse during equilibration with scintillants. This problem has been circumvented by equilibration of the gel with 2,5-diphenyloxazole (PPO) prior to electrophoresis. The presence of PPO in the gel during electrophoresis does not alter mobility of the glycopeptides and oligosaccharides. After electrophoresis, the gel is simply dried and fluorography performed. This allows sensitive and precise comparisons of labeled samples in parallel lanes of a slab gel and, since mobilities are highly reproducible, between different gels. The procedure is preparative in that after fluorography the gel bands can be quantitatively eluted for further study, without any apparent modification by the procedure. In this report, the procedure is illustrated by fractionation of both neutral and anionic glycopeptides produced by the cellular slime mold Dictyostelium discoideum.
D001345:Autoradiography; D004023:Dictyostelium / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005656:Fungal Proteins / Q000032:analysis; D006020:Glycopeptides / Q000032:analysis*; D006023:Glycoproteins / Q000032:analysis; D006031:Glycosylation; D008562:Membrane Glycoproteins; D009844:Oligosaccharides / Q000032:analysis*
D005656:Fungal Proteins; D006020:Glycopeptides; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; C025363:mannoproteins