Re: avidin/biotin blocking in house kit

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From:Phyllis Davie <> (by way of histonet)
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  I was interested in your procedure for in-house avidin/biotin blocking.
 I looked in the sigma catalog, and there are several forms and grades of
avidin and streptavidin available.  Do you have a catalog number?  I only
saw 1 entry for d-biotin, cat# B4501.  Is that the one you use?
     Thanks for your help.

>avidin D 0.01% in PBS, available from Sigma
>d-biotin 0.005% in PBS, available from Sigma
>You could probably add sodium azide 10 - 15 mM to the PBS to preserve the
>avidin solution 15 - 20 min
>rinse well
>biotin solution 15 - 20 min
>rinse well
>Times for avidin and biotin block and rinses can be extended if desired,
>and dependent on the IHC method (seeing differences in some protocols)
>Gayle Callis
>Veterinary Molecular Biology
>Montana State University
>Bozeman MT 59717-3610
>406 994-4705
>406 994-4303

Phyllis Davie
Clinical Laboratory Supervisor
PhenoPath Laboratories
3000 First Ave., #1
Seattle, WA 98121
(206)374-9009  phone
(206)374-9009  fax

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