Detection of Glycosylated Proteins !

iwilson at molbiol.ox.ac.uk iwilson at molbiol.ox.ac.uk
Sun Feb 13 09:01:22 EST 1994


In article <MARDER.147.2D574DF3 at agri.huji.ac.il>, MARDER at agri.huji.ac.il (Jonathan B. Marder) writes:
> In article <2j0boj$7or at mserv1.dl.ac.uk> jawahar at nii.ernet.in writes:
>>Subject: Detection of Glycosylated Proteins !
>>From: jawahar at nii.ernet.in
>>Date: 5 Feb 1994 14:50:59 -0000
> 
>>Hello,
>>Does any one have a method for detecting glycosylated proteins
>>on nitrocellulose membrane by any other method other than silver
>>staining..
>>I am currently using Schiff's reagent but the levels of detections
>>are very low.. Can anyone suggest an improvement over this too !
>>Please send you replies and suggestions to the email address given
>>below..
>>Thank you in advance.
>>jawahar
> You could try probing with biotinylated lectins and then an avidin-lined
> enzyme (which is the same as what you use with biotinylated second antibodies).
> The reagents are reasonably priced and readily available (e.g. from Sigma).
> You should probably try several lectins since they vary in their sugar
> specificities.  I hope that thsi helps.
> __
> Jonathan B. Marder                 '
> Department of Agricultural Botany  |     Internet: MARDER at AGRI.HUJI.AC.IL
> The Hebrew University of Jerusalem | /\/ Bitnet:   MARDER at HUJIAGRI
> Faculty of Agriculture             |/  \ Phone:    (08 or +9728) 481918
> P.O.Box 12, Rehovot 76100, ISRAEL  /     Fax:      (08 or +9728) 467763

Oxford GlycoSystems and Boehringer Mannheim both sell kits for detection of 
glycoproteins, but I've never used them. There's no doubt that Schiff staining 
is NOT the method to use. Also there are a couple of books coming out on 
methods relating to glycosylation. "Glycobiology" one of the Practical 
Approach series is out on 17th Feb. from IRL/OUP (so I'm informed); and the 
Lennarz/Hart edited Methods in Enzymology on similar methods should also be 
coming out. At the moment the older "Carbohydrate Analysis" Practical Approach 
book may have some methodologies of relevance.

Iain Wilson
Dyson Perrins Laboratory, Oxford, UK
iwilson at molbiol.ox.ac.uk



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