SDS of Glycoproteins

labware at crl.com labware at crl.com
Sun Jun 2 17:18:49 EST 1996

p9001808 at vmsuser.acsu.unsw.edu.au wrote:

>We have been trying to identify a membrane bound glycoprotein from a protozoan
>for some months now. We run the crude membrane extract on SDS PAGE 12% (4% 
>stacking gel) and electroblot to PVDF membrane.
>  By carbohydrate staining we have discovered that the glycoprotein won't
>even go into the stacking gel and further, will not transfer to the membrane.
>We have tried several brands of PVDF to no avail. 
>   We have investigated the possibility that we have not properly solubilized
>the membrane (all samples incubated at 95 degrees C for 6 min 1% SDS 800mM 
>2-b-mercaptoethanol and 100mM 2-b-mercaptoethanol included in the PAGE running
>buffer) but this has not helped. Does anyone have any ideas or suggestions?

>                                        David Cross
>                                        University of NSW Australia
>                                        p9001808 at vmsuser.acsu.UNSW.edu.au

How about a phenolic group that is binding? Try urea as h-bond
Other ideas: try some acetone in water-buffer.

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