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PVP-40 for blocking? - Instead of milk (Jan/28/2010 )

I don't seem to get my Western blotting working at all... I have way too much background, horrible unspecific bindings to my ladder and really not constant results in general. Someone recently suggested not using milk for blocking and another one suggested PVP40. So have anyone used Poly vinyl pyrrolidone for blocking and simple did it work out? The article I recently read told to use 0.05% tween 20 and 1% PVP-40 buffer, but am I being overly optimistic :P Even at this point, I'm willing to try anything :D


Have you tried other blockers such as gelatin, BSA, collagen, casein fractions...?


I never did. This PVP-40 was something my boss had read about and what he just told me to do... I modified the entire Western Blotting protocol and surprisingly it started working with 5% milk :lol: So the problem is solved... and we have a large bottle of PVP-40 :D