A method for the identification of sulfated glycopeptide by two-dimensional electrophoresis on cellulose acetate membrane.
Two-dimensional electrophoresis on cellulose acetate membrane permits the clean separation of sulfated glycopeptide in a mixture of acidic glycans (glycosaminoglycans and acidic glycopeptides). Two systems were used. In system 1, 0.1 M pyridine-0.47 M formic acid buffer (pH 3.0) was used in the first and 0.1 M barium acetate (pH 8.0) in the second dimension. In system 2, 0.1 M pyridine-0.47 M formic acid buffer (pH 3.0) was used in the first and 0.1 M HCl in the second dimension. All of the acidic glycans on electrophoretogram were stained with alcian blue in 70% ethanol. On the other hand, sulfated glycans alone were made visible with alcian blue in 0.1 M HCl. Alcian blue in 70% ethanol or 0.1 M HCl, when combined with periodic acid-Schiff's reagent identified sulfated glycopeptides on cellulose acetate membrane.
Yokoyama
M
M
Tanaka
H
H
Yoshihara
S
S
Endo
M
M
D004586:Electrophoresis / Q000379:methods*; D004588:Electrophoresis, Cellulose Acetate / Q000379:methods*; D006020:Glycopeptides / Q000302:isolation & purification; D006025:Glycosaminoglycans / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010189:Pancreatic Juice / Q000032:analysis; D011134:Polysaccharides / Q000302:isolation & purification; D012795:Sialoglycoproteins / Q000302:isolation & purification*
D006020:Glycopeptides; D006025:Glycosaminoglycans; D011134:Polysaccharides; D012795:Sialoglycoproteins; C009354:sulglicotide