Richard,

WE too use the Novocastra antibody.  We were doing it
overnight at 1-100 but recently we have switched to
using Dako TRS (steaming for 30 minutes) then staining
in the CSA Kit from Dako.  The staining is crisper and
more defined than the overnight method which was more
diffuse and "fuzzy".  I also agree that this is one of
the more difficult antibodies and not for the faint of
heart....come on Dako ...give us something better.

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