PAGE of Glycoproteins

p9001808 at vmsuser.acsu.unsw.edu.au p9001808 at vmsuser.acsu.unsw.edu.au
Fri May 31 03:06:36 EST 1996


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



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