Isolation and recovery of acidic oligosaccharides from polyacrylamide gels by semi‐dry electrotransfer
作者:
Ali Al‐Hakim,
Robert J. Linhardt,
期刊:
ELECTROPHORESIS
(WILEY Available online 1990)
卷期:
Volume 11,
issue 1
页码: 23-28
ISSN:0173-0835
年代: 1990
DOI:10.1002/elps.1150110106
出版商: Wiley Subscription Services, Inc., A Wiley Company
数据来源: WILEY
摘要:
AbstractAcidic oligosaccharides derived from glycosaminoglycan heparin were separated by polyacrylamide gradient gel electrophoresis (PAGE). The gel could be visualized using Alcian Blue dye to give a pattern of highly resolved, well defined bands. The particular banding pattern obtained was the result of a heparinase catalyzed depolymerization which afforded oligosaccharide products that differed in size by one disaccharide unit. The separated oligosaccharides could be recovered prior to staining by electroelution onto a positively charged nylon membrane by a semi‐dry transfer procedure. Subsequent elution and quantitative recovery of individual oligosaccharides from the membrane was achieved. By using multiple membrane layers a second separation dimension was obtained, resulting in increased oligosaccharide purity proportional to transfer depth. Preparative gradient polyacrylamide gel electrophoresis followed by semi‐dry electro‐transfer and recovery represents a novel method for the preparation of homogeneous acidic oligosaccha
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