[Histonet] negative immunohistochemistry control

[Sebree Linda A.]

CAP states that for panels, running one negative control using the harshest protocol suffices.

Linda Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-3224
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX: (608)262-7174


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Chris Pomajzl
Sent: Wednesday, March 08, 2006 8:21 AM
To: HISTONET; Rene J Buesa
Subject: Re: [Histonet] negative immunohistochemistry control


I don't understand the idea of only one negative control per case, as
opposed to each antibody. How does one evaluate non-specific staining of
antibodies that do not have a negative control? What if non-specific
staining is inherent to a certain antibody? For example, the new CAP
question regarding staining of endogenous biotin, particularly in kidney and
liver - if you do not run a negative control for that specific antibody, how
do you know if there is non-specific staining?

----- Original Message ----- 
From: "Rene J Buesa" <rjbuesa <@t> yahoo.com>
To: "cynthia haynes" <naje1972 <@t> yahoo.com>;
<Histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, March 08, 2006 7:51 AM
Subject: Re: [Histonet] negative immunohistochemistry control


> Cynthia:
>   The idea of the negative control is to try to determine if any reaction
detected in the case is due to specific or unspecific binding.
>   You don't need to run a negative slide for each antibody you are going
to test on the case, you only need a negative slide per case, no matter how
many antibodies you run. The only requirement is that the negative slide has
to come from the same tissue tested.
>
>   What I used to do was to add buffer instead of the primary Ab and all
the other reagents the same as in the slides run for specific Abs. You could
also add non-immune globuline instead of the antibody but in this case you
would need 1 negative control for each type Ab (1 negative for monoclonal
Abs and 1 for polyclonal Abs).
>   I hope this will help you.
>   René J.
>
> cynthia haynes <naje1972 <@t> yahoo.com> wrote:
>   Good Morning everyone, I have a question about immuno
> staining. I've been away from this type of staining
> for a while and now I am doing them again on a regular
> basis. Why do you run a negative control with each
> run? Are you only suppose to put the normal serum on
> negative control only? I've forgotten; would someone
> please answer these questions for me. Thanks in
> advance.
>
> Cynthia Haynes H.T.
>
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