protocol for removing carbohydrates from cell membranes
mjg177 at psu.edu
Tue Jun 17 22:15:41 EST 1997
Does anyone know of a method for removing the carbohydrates from cell membrane
glycoproteins that will leave viable cells at the end of treatment?
I think periodate oxidation is out of the question. I'm looking for anyone
who may have tried using N or O glycosidases for this purpose. It seems that
most of the literature on the subject deals with protein extracts and not
whole cells. I need whole, viable cells for subsequent cytotoxicity assays.
Secondly, can anyone reommend a universal carbohydrate assay (preferably
colorometric; my fallback position is a gel mobility assay of the membranes
with and without treatment) that I could use to verify carbohydrate release
from my cells into the culture medium supernatant? I've had fairly nice
preliminary results using the anthrone reagent, but it doesn't seem to work on
aminosugars, and will thus probably give a lower than expected reading.
Likewise, some of the other classical tests for reducing sugars might not be
%100 appropriate. Does anyone know if the Molisch reaction will work with
Thanks in advance - Mike
More information about the Methods