Re: [Histonet] IHC background problems on chicken embryos

From:"Katri Tuomala"

I knew you guys would not let me down! Thank you for all excellent replies. 
I will convey these pearls of wisdom to my colleague...


----- Original Message ----- 
From: "Jan Shivers" 
To: "Katri Tuomala" 
Cc: "histonet" 
Sent: Tuesday, September 27, 2005 12:55 PM
Subject: Re: [Histonet] IHC background problems on chicken embryos

>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" 
> To: "HistoNet Server" 
> 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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