Re: nonspecific staining with ventana

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From:amos brooks <atbrooks@snet.net>
To:Vicki L McKaughan <vlm_23@n2mail.com>, histonet <histonet@pathology.swmed.edu>
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Hi,
    Although I don't use a Nexus, I would like to shoot a couple of ideas at you. First have you
tried to reduce the incubation time on the primary antibody? Have you tried titering the antibody
out further (even if it's a predilute)? If you use heat to trim the time down, give it a try
without the heat. Also regardless of what vendors say about their systems not needing protein
blocks, sometimes they help.
Amos Brooks

Vicki L McKaughan wrote:

> We have just purchased the Ventana Nexes Immunostainer.  I am having a problem with heavy
> nonspecific staining on NSE and Melanosome.  I
> think that it is caused by the Amplifier A&B.
> When I remove them from the protocol the stain clears up.  Control tissue stained without the
> amplifier works also.  I was wondering if anyone
> else has run into this problem and what else I
> can try to eliminate it.
>
> Thanks,
> Vicki McKaughan HT (ASCP)
>
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