Vector ABC Elite Kit and biotinylated primary antibodies

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu>
To:histonet@pathology.swmed.edu
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After years of being a purist and non kit user, we have decided to branch
out and work with a universal kit. 

The majority of our primaries (murine CD markers) are biotinylated, and
would like to know any pros and cons on using the ABC Elite kit with these.
 Loss of sensitivity, etc.
We have found the kit to be excellent, particularly with nonbiotinylated
antibodies, but also have a major stock of the others, and happen to like
using them overall.  We have always diluted our own Strepavidin-HRP
substrate, and found this kit to give cleaner results with non-biotin
conjugated primaries.   

My thoughts are if the primary is biotinylated, it should make little
difference, since secondaries are biotinylated also.  As long a dilution
panel is performed with appropriate fixation and blocking, all should work
well. 

What are your experiences?




Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303



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