Western blot for two proteins: any alternative to stripping? - (May/08/2006 )

little off topic, but : for doing a double use membrane, i prepare sample in a 2.2x quantity, and pipett in the same preparation.

I've successfully done a combinatory of first antibody (one was mouse and the other was rabbit) for avoiding stripping, as the membrane was for many blots.

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Problem is that I'm already near to overloading the lanes of the gel (at least to Invitrogen's advice) with tissue whole protein extract because my protein is in it in low concentration...

By the way: Have you ever tried size exclusion chromatography?
Tried it with Sephadex G-75 but it all came down with the first centrifugal spinning.
Was my first try though, so perhaps I'll have another go with less rpm...

And I've never done gel chromatography before so I don't really know what I'm doing

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