Washes for Western, PBS or TBST? - (Mar/13/2006 )

We routinely use PBS with phospho antibodies for both Westerns and cell based assays, and have no difficulties; it's really only an issue if using an alkaline phosphatase secondary. It's very true, though, that using milk as a blocker is a bad idea with phospho antibodies - phosphoepitopes are very abundant in milk.

and by iusing PBS you don't use NaF in it to prevent dephosphorylation of the epitopes?
i was guessing tris was the best for phospho antibody usage.

TBST is ok al well as PBS but be ware Tween 20 could easily wash your proteins away!


cheers,