Re: biotin blocking and non-immune sera for negative control

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From:Debbie Pepperal <>
To:Connie McManus <>
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My 2 peanuts worth. For the biotin block, I do the endogenous block (10
mins in avidin, rinse in PBS, 10 mins in  biotin, rinse in PBS); after the
peroxidase block; before the normal serum incubation.IHC protocol follows

Zenobia Haffajee
HAPS, Newcastle, Australia

 At 02:45 PM 23/05/00 -0600, you wrote:
>At 08:49 PM 05/20/2000 -0400, Amos Brooks wrote:
>>    The blocking is put on just before the primary antibody then it is
>drained or blown off (not rinsed). 
>REALLLY??!!  I was told to use it before H2O2 and rinse vigorously in
>TRIS-HCl with 0.5% Tween.  We have had very good results with what I've
>been doing.
>Connie McManus
>Veterinary Diagnostics Lab
>Utah State University
>Logan, UT

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