Polyacrylamide-gel electrophoresis and Alcian Blue staining of sulphated glycosaminoglycan oligosaccharides

Mary Cowman, M. F. Slahetka, D. M. Hittner, J. Kim, M. Forino, G. Gadelrab

Research output: Contribution to journalArticle

Abstract

Oligosaccharide fragments of glycosaminoglycans may be separated for rapid analysis by electrophoresis through a 10% polyacrylamide matrix. An extensive ladder-like set of bands is observed for partial testicular hyaluronidase digests of chondroitin 4- or 6-sulphate, and for dermatan sulphate. Co-electrophoresis of purified oligosaccharides has established that the major bands of these patterns represent fragments differing in chain length by one disaccharide unit, with the smallest fragments having the greatest mobility. Additional minor bands, representing heterogeneity in the repeating unit structure, are also observed. There are slight differences in the mobilities of oligosaccharides derived from the three major types of sulphated glycosaminoglycans. Alcian Blue is employed for visualization of the digest fragments. Sample loads of 5-10 μg per band appear optimum. The smallest oligosaccharides which may be stained by this method is the hexasaccharide. After consideration of this effect, a good correlation is found to exist between densitometric scans of the gel-electrophoretic patterns and gel-filtration chromatographic profiles based on uronic acid concentration.

Original languageEnglish (US)
Pages (from-to)707-716
Number of pages10
JournalBiochemical Journal
Volume221
Issue number3
StatePublished - 1984

Fingerprint

A73025
Alcian Blue
Electrophoresis
Oligosaccharides
Polyacrylamide Gel Electrophoresis
Staining and Labeling
Gels
Chondroitin
Uronic Acids
Dermatan Sulfate
Hyaluronoglucosaminidase
Disaccharides
Ladders
Glycosaminoglycans
Chain length
Sulfates
Gel Chromatography
Visualization
polyacrylamide gels

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cowman, M., Slahetka, M. F., Hittner, D. M., Kim, J., Forino, M., & Gadelrab, G. (1984). Polyacrylamide-gel electrophoresis and Alcian Blue staining of sulphated glycosaminoglycan oligosaccharides. Biochemical Journal, 221(3), 707-716.

Polyacrylamide-gel electrophoresis and Alcian Blue staining of sulphated glycosaminoglycan oligosaccharides. / Cowman, Mary; Slahetka, M. F.; Hittner, D. M.; Kim, J.; Forino, M.; Gadelrab, G.

In: Biochemical Journal, Vol. 221, No. 3, 1984, p. 707-716.

Research output: Contribution to journalArticle

Cowman, M, Slahetka, MF, Hittner, DM, Kim, J, Forino, M & Gadelrab, G 1984, 'Polyacrylamide-gel electrophoresis and Alcian Blue staining of sulphated glycosaminoglycan oligosaccharides', Biochemical Journal, vol. 221, no. 3, pp. 707-716.
Cowman, Mary ; Slahetka, M. F. ; Hittner, D. M. ; Kim, J. ; Forino, M. ; Gadelrab, G. / Polyacrylamide-gel electrophoresis and Alcian Blue staining of sulphated glycosaminoglycan oligosaccharides. In: Biochemical Journal. 1984 ; Vol. 221, No. 3. pp. 707-716.
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