title: Direct detection of beta-1,3-glucanase isozymes on polyacrylamide electrophoresis and isoelectrofocusing gels.
abstract: A procedure to assay isozymes of beta-1,3-glucanase directly on polyacrylamide gel electrophoresis (PAGE) and isoelectrofocusing (IEF) gels by using 2,3,5-triphenyltetrazolium chloride is described. The reagent reacts with reducing sugars released by beta-1,3-glucanases from the substrate laminarin. Acidic and neutral isozymes of beta-1,3-glucanase were detected and quantified on 17.5% native PAGE gels run with an anodic buffer system. A significant linear relationship (alpha = less than 0.01, R = 0.991) was observed between amounts of beta-1,3-glucanase loaded and intensity of bands stained with the reagent on native PAGE gels. A full isozyme pattern was obtained on 7.5% IEF gels with a pH range of 3.5-9.5. The IEF gels were heated in a microwave oven during the staining process to minimize diffusion.
authors: Department of Plant Pathology, University of Kentucky, Lexington 40546.
authors: S Q
authors: SQ
authors: Pan
authors: X S
authors: XS
authors: Ye
authors: J
authors: J
authors: Kuć
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D043326:Glucan 1,3-beta-Glucosidase; D005959:Glucosidases / Q000032:analysis*; D007525:Isoelectric Focusing / Q000379:methods; D007527:Isoenzymes / Q000032:analysis*; D013194:Staining and Labeling; D013778:Tetrazolium Salts; D001617:beta-Glucosidase / Q000032:analysis*
chemical_list: D007527:Isoenzymes; D013778:Tetrazolium Salts; C009591:triphenyltetrazolium; D005959:Glucosidases; D001617:beta-Glucosidase; D043326:Glucan 1,3-beta-Glucosidase