Quantitation of Sugars on glycoproteins

Steven Pirie-Shepherd srps at galactose.mc.duke.edu
Wed May 10 07:08:30 EST 1995


Frank C Hay (frank at rabbits.demon.co.uk) wrote:
: 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.

Maybe a modification of the Boehringer kit?

Boehringer use dig-labelled lectins and an anti-dig antibody linked to 
AP. What about dig labelled lectins, and hot anti-dig antibody??

Count by using either a phosphor Imager or cutting out the bands from the 
inert support and counting in a gamma counter? Express results as cpm per 
ug protein? Use fetuin or some such as a standard and construct a 
standard curve.


This is juat an idea, not a solid proposal as yet.

__________________________________________________
Steven Pirie-Shepherd		       		
srps at galactose.mc.duke.edu	        
"Insert your own pithy phrase just about here!"



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