[Histonet] Help!!!

[vsailes <@t> nd.edu]

Hello Histonetters,
I'm working on an IHC protocol for Albumin -- staining FFPE mouse tissues.  I'm
new to working out conditons for antibody protocols and could use some help.I'm
testing the antibody at different dilutions and I appear to have positive
staining but I'm also seeing small areas in the negative that shows positive
staining. Liver is my control and I use steamer (HIER) for 30 minutes, my steps
include a 30 min quench in 1.6% hydrogen peroxide, NS block, Antibody, Secondary
antibody(Biotin), Streptavidin, Chromogen, Counterstain
My concern is that I'm using buffers that contain BSA and I'm wondering if that
is causing staining to appear or should I be using avidin/biotin block along
with a peroxide block. Any suggestion would be helpful. Thank you for your help
in advance.
Valerie