Re: Negative Reagent Controls For Immuno

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From:Bonnie P Whitaker <>
To:histonet <>
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Hi Amos,

I disagree.  The purpose of the negative control is (at least in my mind) a
control on endogenous activity in each tissue and non-specific binding.  For
example if you are using an avidin-biotin method, then the negative control
would indicate any endogenous biotin activity that had not been blocked,
thus preventing confusion by whomever is interpreting the results.  The same
for other tissue components that pick up non-specific staining..... if your
negative has positivity, then it can be "mentally subtracted" or in a severe
case, rerun using different blocks, or techniques,etc.

Bonnie Whitaker

----- Original Message -----
From: amos brooks <>
To: Mackinnon, John <>
Cc: HistoNet (E-mail) <>
Sent: Saturday, February 26, 2000 8:45 AM
Subject: Re: Negative Reagent Controls For Immuno

> Hi,
>     I see no use running a negative control for each block if the
processes are
> identical. Exception being the Herceptest, solely because it is FDA
> and that is what they expect of this method. It is appropriate to have one
> representative control but it needn't be one for each block. That would be
> redundant, impractical, and unnecessary. We've never had a deficiency for
> and I would like to hear of anyone who has.
> Amos Brooks
> "Mackinnon, John" wrote:
> > When running negative reagent controls you should be running a negative
> > each block, if you are running both monoclonal and polyclonal antibodies
> > should run a negative for each of these.  If one of your antibodies
> > HIER another enzyme and one no pretreatment you should be running a
> > for each of these.  All of your controls must be treated the same way
> > your test sections are treated, if you don't do this you are not really
> > controlling the procedure.  Expense is no excuse for not running proper
> > I can't remember ever hearing a biochemist saying that they were not
> > to run all of the appropriate QC because it seemed excessive and
> >
> > John MacKinnon MLT, ART
> > Senior Technologist, Pathology
> > Lakeridge Health Oshawa
> > Ontario, Canada

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