[Histonet] Negative control for immuno slides

[Bryan Hewlett]

Anissa,

The ideal way of running negative reagent controls is to substitute an Ig
isotype-matched Monoclonal antibody, directed against an antigen not
normally found in human material, at the same concentration as your primary
antibody in the same diluent.
Alternately, including in your panel of antibodies another monoclonal
primary antibody of the same Ig isotype, using the same pre-treatment but
with a totally different distribution of reactivity, is acceptable as a
control. Failing that, simply applying the primary antibody diluent, as you
are doing, is acceptable according to NCCLS guidelines.
For each case, following each method variation, the negative reagent control
for each primary must be applied as you are doing.
However, should you use several monoclonal antibodies on a case, of the same
Ig isotype with the same pre-treatment,  then only one negative reagent
control for those would be required.

Bryan Hewlett
Technical Consultant
QMP-LS,
Ontario, Canada


----- Original Message -----
From: "Anissa Choi" <anissachoi <@t> hotmail.com>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Friday, May 07, 2004 12:22 AM
Subject: [Histonet] Negative control for immuno slides


> We do our negative reagent control by omitting the primary antibody. The
> number of negative
> slides we run depends on the pre-treatment of the antibodies requested on
> the case. We run one
> negative slide for each method i.e. no pre-treatment, protease and
pressure
> cook. Are we doing
> more negative control slides than what is required ? I know some people
just
> do one negative
> slide per case. Any comments on the proper way of running negative immuno
> controls? Thanks
> in advance for your inputs !
>
>
> Anissa Choi
> Burnaby Hospital
> Burnaby, B.C.
> Canada
>
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