Re: Vector ABC Elite Kit and biotinylated primary antibodies

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From:Luis Chiriboga <>
To:Gayle Callis <>
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The advantage of using a biotinylated secondary antibody is that you increase
the signal to noise ratio. You may lose some sensitivity but the improved S/N
will give better contrast . Vector kits are excellent and after using their
products for several years,  I would recommend them.  I have used biotinylated
and FITC labeled primaries with Vector and Ventana detection systems without to
much problem.  It is essential that the proper controls be used when
determining the optimal conditions for any antibody assay
good luck

Gayle Callis wrote:

> 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

Luis Chiriboga Ph.D HT(ASCP)QIHC
Department of Pathology 4W27
Bellevue Hospital
NYU School of Medicine
First Avenue & 27th Street
New York, NY 10016

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