I have a doubt about native PAGE gel preparation and will greatly apreciate any help from
you "protein expert" guys :-)
I am doing 7% native acrylamide gels to separate two isoforms of glutamine synthetase (GS)
from plant crude extracts and then stain the activity in-gel before coomassie. My problem is that
one isoform is unstable and inactivates during the electrophoresis (I normally run it overnight
in a cold room) so I am looking for some way to improve its stability during the run.
As the enzyme in the crude extracts is stabilised by DTT, glycerol and glutamate (that is a GS substrate)
I was thinking if is it possible to add this compounds in the gel! Does anyone knows if this compounds
can be added to the native PAGE gel and /or loading buffer???
Thanks a lot for your help,
Marco Betti Ph.D.
Departamento de Bioquímica vegetal y Biología Molecular
Universidad de Sevilla, Spain