> Here is what I concluded about dots problem for myself:
>> Dots are result of small pieces of gel sticking to the
> membrane. Antibody can get trapped in the gel, and then are
> washed out poorly (diffusional problems), resulting in
> intense localized signal upon development with low MW
> substrates. That's why most of the dots are usually
> located on the periphery of the blot - where gels
> tend to stick most.
>> Now I inspect blots visually and try to remove any
> gel pieces with a jet from squeeze bottle. I think
> it helps.
>> - Dima
In my case it was bubbles sticking persistantly to the membrane. I had to
use a gloved finder to poke and squash them off the membrane during
blotting and washing. Mike.
