[Histonet] negative immunohistochemistry control

[Luck, Greg D.]

Chris,

The CAP question is really designed to address the integrity of your detection system not the antibodies in general.  If your negative control is truly negative (as it should be) any staing on the positive patient slide should be attributable to the antibody reagent that was applied as opposed to something unique to the tissue (excluding some tissue artifacts that seem to stain with many antibodies, like necrotic tissue components for example).

Greg Luck, BS, HT(ASCP)
Anatomic Pathology Supervisor
Deaconess Medical Center
800 W. 5th Ave
Spokane, WA 99204
Phone 509.473.7077
Fax 509.473.7133
luckg <@t> empirehealth.org
www.deaconessmedicalcenter.org


-----Original Message-----
From: Chris Pomajzl [mailto:cpomajzl <@t> cpllabs.com] 
Sent: Wednesday, March 08, 2006 6:21 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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