Re: "negative controls for immunoperoxidase

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From:Victoria Baker <>
To:Mary Latino <>
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Negative controls one is referred to as serum control
(this verifies that your procedure is correct) and
negative tissue control (a tissue that is negative to
the antigen you are testing for, best one, a normal
lung for p53).  In the clinical setting usually only a
negative serum control is used.  You do need to run a
negative control slide for mouse or rabbit.  Along
with a control tissue negative.  If for one case you
have 3 blocks with 4 different antibodies, 2 mouse 2
rabbit.  I usually would pick a representative block
from each species and use it as a negative.  After all
if your doing the same procedure for all the same
blocks in the same run, why run individual blocks.  I
was never questioned by CAP about this either and the
lab always passed with no deficiencies.

Good luck!

Vikki Baker
American Health Foundation
Valhalla, New York


--- Mary Latino <> wrote:
> I am curious as to what everyone is doing for their
> "negative controls" for immunohistochemistry (IHC). 
> The current CAP check list, basically says that you
> need a negative control slide for every Ab that you
> use for each block. This seems ridiculous to me, but
> only because of the way we have set up our IHC
> procedures. Presently we buy prepared negative
> control
> sera for rabbit and mouse Abs from Biogenex.  This
> is
> probably not the best way to stain negative
> controls.
> Presently we are manually staining with the DAKO
> Envision+ system and use concentrated Abs.
> We are also looking into getting an automated
> stainer
> so the recent discussion has been a big help to us.
> Mary Latino
> Pathologist's Assistant
> Nyack Hospital
> Nyack, NY 10960  
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