title: Quantitative determination of sulfated glycopeptide by two-dimensional electrophoresis on cellulose acetate membrane.
abstract: Quantitative determination of the sulfated glycoproteins present in tissue and secretion fluid was performed. After digestion of the specimen with pronase in order to convert glycoproteins to glycopeptides, the sulfated glycopeptides were separated from a mixture of acidic glycans (glycosaminoglycans, sialoglycopeptides and sulfated glycopeptides) by two-dimensional electrophoresis on cellulose acetate membrane [(1986) J. Biochem. Biophys. Methods 12, 239-246]. After staining with alcian blue, the spot of sulfated glycopeptide on the cellulose acetate membrane was cut out, and then only the dye bound to the sulfated glycopeptide was extracted with a 5% cetylpyridinium chloride solution at 100 degrees C for 15 min. The extract was then measured by absorbance at 615 nm using an authentic sulfated glycopeptide as a standard. This method facilitated the determination of sulfated glycopeptides, which were separated from other acidic glycans, within the range 0-25 micrograms.
authors: Department of Biochemistry, Hirosaki University School of Medicine, Japan.
authors: K
authors: K
authors: Muramoto
authors: H
authors: H
authors: Tanaka
authors: S
authors: S
authors: Kimura
authors: K
authors: K
authors: Kubo
authors: S
authors: S
authors: Yoshihara
authors: M
authors: M
authors: Yokoyama
authors: Y
authors: Y
authors: Yoshida
authors: M
authors: M
authors: Endo
mesh_terms: D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D051152:Clusterin; D004588:Electrophoresis, Cellulose Acetate / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006023:Glycoproteins; D006025:Glycosaminoglycans; D007202:Indicators and Reagents; D007413:Intestinal Mucosa / Q000032:analysis; D007421:Intestine, Small / Q000032:analysis; D018832:Molecular Chaperones; D011402:Pronase; D013552:Swine
chemical_list: D002241:Carbohydrates; D051152:Clusterin; D006023:Glycoproteins; D006025:Glycosaminoglycans; D007202:Indicators and Reagents; D018832:Molecular Chaperones; D011402:Pronase