biotin non-specific binding

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From:Hugo <ivha23@blues.uab.es> (by way of histonet)
To:histonet@histosearch.com
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Hi to all!
I wonder if someone out there can help me on this. I routinely do IHC's
on bird tissue (ranging from sparrows to toucans to poultry and so on).
I've always had trouble with avidin-biotin methods (excessive
non-specific staining, NOT background). Lately I've come to the
conclusion that my trouble was non-specific biotin-binding sites in the
tissue. Using a commercial biotin blocking reagent (apparently it's just
a biotin solution) at 4x recommended concentration has practically
eliminated all the non-specific staining.
My questions are:
Anyone else out there work with bird tissue and had similar problems?
Do you think I could just buy biotin, make a solution in TBS (or
whichever buffer) and apply it as a blocker?

Thanks

Hugo Fernandez Bellon
U.D. Histologia i Anatomia Patologica
Fac. de Veterin`ria, UAB
SPAIN
ivha23@blues.uab.es




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