Native gel

Shanthi Adimoolam shanth at ux7.cso.uiuc.edu
Tue Apr 16 09:01:05 EST 1996


Hi,
I am trying to run a native gel to see if two fragments from a trypsin 
digest are in some way covalently associated or not. But for some reason, 
the intact protein (undigested) runs very very close to the dye front, 
and the other smaller proteins are stacked at the dye front. Hence i am 
not able to see the tryptic fragments. The running dye I use is .01% BPB 
in 10mM tris, 1mM EDTA, pH8.0. The gel i ran was a 12.5% homogenous gel 
and the intact protein that i digested is ~67kD and it runs almost at the 
dye front.

I need to do this pretty urgently, any suggestions would be welcome.

Thank you very much
-shanthi




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