Protocol Online logo
Top : New Forum Archives (2009-): : Electrophoresis

SDS-PAGE running conditions - (Nov/14/2011 )

Hey guys,

I'm in a bit of a dilemma currently.

I want to push my proteins as tightly as i can within my stack to the resolving without much spread.
On one hand i can run it at 150V for 5-10mins to get everything through the stack and into the resolving
and on the other hand i can run it at 30V for 2-3hours to get the same result as above.

These would then be cranked up/down to say 60-90V to allow for the proteins to run through the resolving gel.

My question is whether the slower method is better than the faster method, or if anyone has any other ideas to get nice tight bands.

(These are all being run at 4degC)



Technically both will give the same result, as the separation is dependent on the percentage of the gel, not the voltage.