Re: [Histonet] Using Cy2 immunofluorescence?

From:Gayle Callis

For a biotinylated primary, you should come back with Strepavidin 
conjugated to the CY2 fluorophore.  I don't think the secondary conjugated 
to Cy2 will like all that biotin in the way when you try to connect primary 
to secondary.   Jackson Immunoresearch sells SA-Cy2, not terribly expensive 
and they have excellent products.

If you use a biotinylated primary, be sure to use an avidin/biotin block or 
better yet, Strepavidin/biotin blocking kit (both available from Vector).


At 09:02 AM 8/26/2004, you wrote:
>We are using a cy2 immunofluorescent secondary antibody for IHC, and
>we would like to know if there are any complications using it on a
>biotin conjugated primary. We are not sure if there will be
>interferance with the signal or if there will be no sites on the
>primary ab for the secondary to bind. I appreciate your help.
>Peter Fanti
>University at Buffalo
>OB/GYN department
>pcfanti@buffalo.edu
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)



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