Determination of the concentrations of oligosaccharides, complex type carbohydrates, and glycoproteins using the phenol-sulfuric acid method.
The concentrations of methyl glycosides, oligosaccharides, glycopeptides, and glycoproteins can be accurately determined by using calibration curves composed of the appropriate monosaccharide(s) obtained with a modified version of the colorimetric phenol-sulfuric acid method. Calibration curves of micrograms sugar vs. 490 nm for Man, Glc, or Gal are shown to provide reliable determinations (typically +/- 3-4%) of corresponding methyl glycosides and linear and branched-chain oligosaccharides containing the corresponding reactive hexose residue. For complex oligosaccharides containing a known mixture of reactive hexose units, the appropriate mixture of monosaccharides are shown to provide equally accurate calibration curves for concentration determinations. In the case of the soybean agglutinin, which is a tetramer possessing one Man9 oligomannose-type chain per subunit, the protein concentration was determined from the Man calibration curve which agreed with that obtained from the molar extinction coefficient of the protein.
Saha
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Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY 10461.
Brewer
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D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D003124:Colorimetry / Q000379:methods; D005643:Fucose; D005690:Galactose; D005947:Glucose; D006020:Glycopeptides / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis*; D006027:Glycosides / Q000032:analysis; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy; D008358:Mannose; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry*; D019800:Phenol; D010636:Phenols; D011134:Polysaccharides / Q000032:analysis*; D013329:Structure-Activity Relationship; D013464:Sulfuric Acids
D006020:Glycopeptides; D006023:Glycoproteins; D006027:Glycosides; D007202:Indicators and Reagents; D009844:Oligosaccharides; D010636:Phenols; D011134:Polysaccharides; D013464:Sulfuric Acids; D005643:Fucose; D019800:Phenol; D005947:Glucose; C033158:sulfuric acid; D008358:Mannose; D005690:Galactose