Hope someone could help me.
I’m trying SDS-PAGE for visualisation of polysaccahrides, and I wanted to try different % of polyacrylamide.
Working with protein, I’ usually use 12% gels, in a continuous system buffer (Laemmli buffer), and I use 20mA/gel.
To separate polysaccharide, the porotocole I follow (and bibliography) recommend a discontinuous system buffer, and it recommends also to run at constant voltage (so the protein electrophoresis guide I’m using to refresh concepts).
In the very fist paper that evaluate the use of this discontinuous buffer system for polysaccharides, the SDS-PAGE conditions were 16%, 30V while sample were passing through the stacking gel, then 100V when samples entered the separating one.
I’ve tried this conditions with a 12% gel, and the profile was very nice…, but I’m ready to try a 9% gel, and I hesitate in choosing the voltage…