Western Blotting loading control for large proteins - (Apr/30/2013 )

Hi,

For some time I have had a problem with loading contol for my protein, which is ~250kDa.

Running SDS-page gel, I usually run it so long (5% gel) that anything below 170kDa escape my gel and I can't use any of loading control like GAPDH, Actin, tubulin. I work with HeLa/Hek293 T cells.

I know that I can run a separate gel, run it not so long just to make a loading control (detection of protein ~<100kDa) but I would prefer to have a control on the same gel, it is more reliable.

Any suggestions?


Alchemic

run a gradient gel.

Thanks for prompt reply. I will try that.