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Discontinuous/Continuous SDS PAGE - (Feb/09/2005 )

When do you use Discontinuous and when Continuous SDS PAGE? Do I always have to use a stacking and a resolving gel?



Of course you have to use both a stacking AND a resolving gel. The stacking gel will "stack" the sample you loaded in a sharp band, which will the be separated in the resolving gel, according to their MW. Without the stacking gel, your sample would separate as a smear and each protein could appear as thick as the well height...

The use of a continuous gel (increasing in acrylamide concentration) is, for me, just useful if you want to resolve two (or more) proteins very close in mass.

Hope I could help..