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Quantitative determination of sulfated glycopeptide by two-dimensional electrophoresis on cellulose acetate membrane.
MedLine Citation:
PMID:  2809069     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Quantitative determination of the sulfated glycoproteins present in tissue and secretion fluid was performed. After digestion of the specimen with pronase in order to convert glycoproteins to glycopeptides, the sulfated glycopeptides were separated from a mixture of acidic glycans (glycosaminoglycans, sialoglycopeptides and sulfated glycopeptides) by two-dimensional electrophoresis on cellulose acetate membrane [(1986) J. Biochem. Biophys. Methods 12, 239-246]. After staining with alcian blue, the spot of sulfated glycopeptide on the cellulose acetate membrane was cut out, and then only the dye bound to the sulfated glycopeptide was extracted with a 5% cetylpyridinium chloride solution at 100 degrees C for 15 min. The extract was then measured by absorbance at 615 nm using an authentic sulfated glycopeptide as a standard. This method facilitated the determination of sulfated glycopeptides, which were separated from other acidic glycans, within the range 0-25 micrograms.
Authors:
K Muramoto; H Tanaka; S Kimura; K Kubo; S Yoshihara; M Yokoyama; Y Yoshida; M Endo
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of biochemical and biophysical methods     Volume:  19     ISSN:  0165-022X     ISO Abbreviation:  J. Biochem. Biophys. Methods     Publication Date:  1989 Jul 
Date Detail:
Created Date:  1989-12-12     Completed Date:  1989-12-12     Revised Date:  2005-11-17    
Medline Journal Info:
Nlm Unique ID:  7907378     Medline TA:  J Biochem Biophys Methods     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  75-82     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, Hirosaki University School of Medicine, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Carbohydrates / analysis
Clusterin
Electrophoresis, Cellulose Acetate / methods
Electrophoresis, Gel, Two-Dimensional / methods
Glycoproteins*
Glycosaminoglycans*
Indicators and Reagents
Intestinal Mucosa / analysis
Intestine, Small / analysis
Molecular Chaperones*
Pronase
Swine
Chemical
Reg. No./Substance:
0/Carbohydrates; 0/Clusterin; 0/Glycoproteins; 0/Glycosaminoglycans; 0/Indicators and Reagents; 0/Molecular Chaperones; EC 3.4.24.-/Pronase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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