[Histonet] IHC background problems on chicken embryos

[Jan Shivers]

I would second Brian Chelack's suggestion to use a NON-biotinylated IHC
staining system with avian tissue.  I use EnVision+/HRP exclusively (I stain
for a few avian viruses here).  I would also suggest adding 2% normal
chicken serum per volume to the EnVision+/HRP reagent.  It helps block
nonspecific binding of the EnVision to endogenous chicken Igs and serum
components.

Jan Shivers
U of MN Vet Diag Lab

----- Original Message ----- 
From: "Katri Tuomala" <katri <@t> cogeco.ca>
To: "HistoNet Server" <histonet <@t> lists.utsouthwestern.edu>
Sent: Monday, September 26, 2005 8:09 PM
Subject: [Histonet] IHC background problems on chicken embryos


> Hi Histonetters,
> A colleague of mine is trying to get VWF (Dakocytomation) and VEGF(R&D
> Systems) working on chicken tissue. The antibodies are performing well on
> human and rat tissue, but excessive background is the problem with the
> chicken. Would the blocking serums (normal goat and normal rabbit
> respectively) be the problem as well as the biotinylated secondary
reagents
> raised in goat and rabbit? My experience is strictly with human tissues,
so
> I am stumped. To my knowledge neither of these antibodies have been proven
> to work with chicken, so maybe that is the problem.
> Any advice from "chicken" people?
>
> Katri
>
> Katri Tuomala
> Hamilton, Ontario, Canada
>
>
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