No retitering needed, I did make sure I added 100 ul to the well, could it be
you are not adding the exact amount needed to flow across the section.

I pipette the exact amount, but if you use dropper bottles, know how much is in
the drop for your kit system, and make sure the total number of drops adds up
to 100 ul.  and add when you know the time is up after draining away the
buffer, we do 4 - 5 minutes since the plus charge marks (little + signs)
creates bigger gap, and give a bit faster flow, no problems though.

also found too many sections on the slide alters the flow, plus I mount them in
the bottom 1/3 of slide, not close to the label, or offset the sections bit, so
one does not impede the flow to a section below it.  

We never added detergents with our antibodies, but never found they deterred
staining with coverplates.

Gayle Callis