PMID: 8595254Oct 1, 1995Paper

Analysis of serine/threonine-linked oligosaccharides derived by alkaline-borohydride treatment of mucin glycoproteins electroblotted onto membranes: comparison of the saccharide profiles of the 390 kDa and 350 kDa forms of epitectin

Glycoconjugate Journal
Q Zhu, V P Bhavanandan

Abstract

Alkaline borohydride treatment is widely used for the release of carbohydrate moieties from O-glycosylated glycoproteins and mucins. We have adapted this procedure to micro quantities of glycoproteins blotted on membranes. After electrophoresis and transfer to nitrocellulose, nylon or polyvinylidene difluoride membrane, alkaline borohydride treatment was done directly on glycoprotein containing areas of membrane which were cut out with the aid of guide strips stained with Coomassie Blue or lectin-digoxigenin. In combination with standard saccharide fractionation techniques, this procedure can be used to characterize the oligosaccharides of mucins or mucin-type glycoproteins that are separated by gel electrophoresis from crude sources. Using this approach we have characterized the saccharides derived from the two species of epitectin, a malignancy-associated mucin type glycoprotein, isolated from metabolically labelled H.Ep2 cells.

References

Mar 9, 1976·Biochemistry·H Furthmayr, V T Marchesi
Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·H TowbinJ Gordon
Jan 1, 1977·Journal of Supramolecular Structure·H Furthmayr
Jan 1, 1992·Critical Reviews in Biochemistry and Molecular Biology·G J Strous, J Dekker
Nov 1, 1991·Glycobiology·V P Bhavanandan
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May 15, 1989·Analytical Biochemistry·G I TousS Stein
Aug 1, 1981·Carbohydrate Research·M L BerghD H van den Eijnden

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