Western blot

Don Incognito imperat at hotmail.com
Thu Oct 28 22:13:53 EST 2004


hubahopp at gmx.de (Wolfgang Schechinger) wrote in message news:<3.0.6.32.20041025113622.0101bdd0 at pop.gmx.net>...
> Hm - what about CREB and Phospho-CREB? 
> 
> If you want positive controls, they are most suitable. You might have a
> cell line stably overexpressing CREB, so you could treat one dish with
> vanadate / fluoride for some time before lysing it and another one with a
> suitable phosphatase (CIP/SAP?) after lysing it.

I believe Andrea was asking about LOADING controls, not POSITIVE
controls. Suitable loading controls are legion, and depend on the cell
line you're using. Beta-actin, GAPDH and tubulin all come to mind.

Warning: these are often expressed at very high levels. I would not
use antibodies against them on the same blot as you're using for your
CREB and p-CREB targets.

Hope this helps.

Don Incognito
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