Can one keep NC menbrane overnight and do ECL the next day? - (Jan/07/2007 )

Can one keep a blot (on nitrocellulose) after antibody treatment and subsequent washings overnight in PBS and detect the bands (ECL) the next day?

yes, I think there is no problem. However, what I prefer (although I work with PVDF membranes but I think in this case it doesn´t matter) is incubating overnight 4ºC with the primary antibody (this works fine and reduces unespecific binding) and the next day wash and incubate with the secondary and then ECL.

I agree with Pumuki.
I never tried overnight in wash buffer, however overnight in primary antibody is fine.
I don't see why overnight in wash buffer would be bad.

Some times we would wash the nitocellulose after primary antibody and dry it between the filter paper and complete the western with secondary whenever we have time. Works nicely.

Thanks Fellows!
Actually, my dilemma was unrelated to blocking/nonspecificity. I was already through with the experiment before I realized that I had finished the ECL kit! However, it worked. No problems.