Running different % acyrlamide gels - (Mar/17/2008 )
Hi,
I am going to run two different gels, one at 7.5% (looking for a 150 kDa protein) and one at 12% (looking for a 27 kDa protein). Can these two gels be run together in the same bio-rad set-up? I usually run my samples through the stacking gel at 70V and then increase the voltage to 130V. I just want to be sure that the proteins will run at comparable rates and that their elecrophoresis will end at the same time. Thanks.
U can run diffent percentage of gels together
i am running 12 % and 15 % gels together and i didnt see any difference
I am going to run two different gels, one at 7.5% (looking for a 150 kDa protein) and one at 12% (looking for a 27 kDa protein). Can these two gels be run together in the same bio-rad set-up? I usually run my samples through the stacking gel at 70V and then increase the voltage to 130V. I just want to be sure that the proteins will run at comparable rates and that their elecrophoresis will end at the same time. Thanks.
The only thing that may be different is the time needed to go through a 12% vs a 7%. As I experienced, it takes less time to migrate through a smaller percentage. But since the difference is quite small (less than 10min), everything should go fine.
actually, you should not run two different gels in the same unit or with the same power supply (at the same time).
the current will favor one over the other (path of least resistance). not only will one run faster but will also run hotter than the other.
reproducibility is questionable.
you may be able to get away with it but it is not recommended.