Combined alcian blue and silver staining of glycosaminoglycans in polyacrylamide gels

Application to electrophoretic analysis of molecular weight distribution

Hoki Min, Mary Cowman

Research output: Contribution to journalArticle

Abstract

Oligomeric and polymeric fragments of glycosaminoglycans may be separated for rapid analysis by electrophoresis through a 10% polyacrylamide matrix. A ladder-like series of bands is observed, in which adjacent major bands correspond to species differing in chain length by one disaccharide unit. The component species are detected by a combined alcian blue and silver staining protocol. Detection limits are less than 50 ng per band, or approximately 2-5 μg total load for polydisperse samples. Densitometry of the stained gel may be used to determine molecular weight averages and distribution. The applicable molecular weight ranges are approximately 4000 to 100,000 for hyaluronate, or 1500 to 40,000 for chondroitin and dermatan sulfate samples of moderate charge density heterogeneity.

Original languageEnglish (US)
Pages (from-to)275-285
Number of pages11
JournalAnalytical Biochemistry
Volume155
Issue number2
DOIs
StatePublished - 1986

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Alcian Blue
Silver Staining
Molecular weight distribution
Glycosaminoglycans
Silver
Molecular Weight
Molecular weight
Dermatan Sulfate
Densitometry
Chondroitin Sulfates
Disaccharides
Ladders
Charge density
Electrophoresis
Chain length
Limit of Detection
Gels
polyacrylamide gels
polyacrylamide

Keywords

  • chondroitin sulfate
  • dermatan sulfate
  • electrophoresis
  • glycosaminoglycans
  • hyaluronate
  • staining

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

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title = "Combined alcian blue and silver staining of glycosaminoglycans in polyacrylamide gels: Application to electrophoretic analysis of molecular weight distribution",
abstract = "Oligomeric and polymeric fragments of glycosaminoglycans may be separated for rapid analysis by electrophoresis through a 10{\%} polyacrylamide matrix. A ladder-like series of bands is observed, in which adjacent major bands correspond to species differing in chain length by one disaccharide unit. The component species are detected by a combined alcian blue and silver staining protocol. Detection limits are less than 50 ng per band, or approximately 2-5 μg total load for polydisperse samples. Densitometry of the stained gel may be used to determine molecular weight averages and distribution. The applicable molecular weight ranges are approximately 4000 to 100,000 for hyaluronate, or 1500 to 40,000 for chondroitin and dermatan sulfate samples of moderate charge density heterogeneity.",
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T2 - Application to electrophoretic analysis of molecular weight distribution

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N2 - Oligomeric and polymeric fragments of glycosaminoglycans may be separated for rapid analysis by electrophoresis through a 10% polyacrylamide matrix. A ladder-like series of bands is observed, in which adjacent major bands correspond to species differing in chain length by one disaccharide unit. The component species are detected by a combined alcian blue and silver staining protocol. Detection limits are less than 50 ng per band, or approximately 2-5 μg total load for polydisperse samples. Densitometry of the stained gel may be used to determine molecular weight averages and distribution. The applicable molecular weight ranges are approximately 4000 to 100,000 for hyaluronate, or 1500 to 40,000 for chondroitin and dermatan sulfate samples of moderate charge density heterogeneity.

AB - Oligomeric and polymeric fragments of glycosaminoglycans may be separated for rapid analysis by electrophoresis through a 10% polyacrylamide matrix. A ladder-like series of bands is observed, in which adjacent major bands correspond to species differing in chain length by one disaccharide unit. The component species are detected by a combined alcian blue and silver staining protocol. Detection limits are less than 50 ng per band, or approximately 2-5 μg total load for polydisperse samples. Densitometry of the stained gel may be used to determine molecular weight averages and distribution. The applicable molecular weight ranges are approximately 4000 to 100,000 for hyaluronate, or 1500 to 40,000 for chondroitin and dermatan sulfate samples of moderate charge density heterogeneity.

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