Quantification of Sugars on glycoproteins

John Proudman, GGPL, B-262 johnp at GGPL.ARSUSDA.GOV
Thu May 11 09:05:09 EST 1995


Frank:
You might try the "DIG Glycan ELISA" sold by Boehringer Mannheim (Cat. 
#1460366).  You adsorb your glycoprotein to the wells of an ELISA plate, 
incorporate DIG into the CHO by oxidation, and quantify the DIG relative to a 
fetuin standard.  The standard curve goes down to 2 ng of fetuin.  In my 
limited experience, the standard curve works very well and the sensitivity is 
as advertised.  How well it detects your oligosaccharide may depend on the 
amount of CHO present and perhaps its tertiary structure (accessability of 
the CHO to reagents).

John

John A. Proudman
Germplasm & Gamete Physiology Laboratory
Beltsville Agricultural Research Center
Agricultural Research Service
U.S. Department of Agriculture
Beltsville, MD 20705 USA


>
>Further to my query on inhibitors on glycosidase inhibitors is there is a
>simple way of quantitating total oligosaccharide on  glycoproteins on
>Western blots?
>I know Oxford Glycosystems has a technique for determining whether a
>protein is glycosylated but they tell me this is not very quantitative.
>
>Thanks for all the replies to my previous query which were sent to me
>directly.
>
>-- Frank C Hay
>Division of Immunology
>St George's Hospital Medical School
>London SW17 0RE, UK
>Tel: 0181 725 5751, Fax: 0181725 3549,
> email: frank at rabbits.demon.co.uk



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