There is another reason to use an irrelevant antibody as a negative
control, especially when using ascites fluid.

The normal serum is not a 'hyper-immune' serum, that is, it's host has
not been challenged with a large amount of an antigen, so it will most
likely not have a super-abundance of any single antibody in it. So, it
is much less likely to cause background than a "hyper-immune" serum or
ascites fluid.

Using an "irrelevant" antibody, which has been prepared in the same way
as your antibody of interest, is the best because it will have a
super-abundance of one or more antibodies and is more likely to cause
non-specific background than a normal serum.


Tim Morken, B.A., EMT(MSA), HTL(ASCP)
Infectious Disease Pathology
Centers for Disease Control
MS-G32
1600 Clifton Rd.
Atlanta, GA 30333
USA

email: tim9@cdc.gov
       timcdc@hotmail.com

FAX:  (404)639-3043


----Original Message Follows----
Date: Tue, 02 Mar 1999 14:56:09 -0500
From: Jeff Silverman <peptolab@hamptons.com>
Subject: Re: Irrelevant antiibodies versus omiting the primary Ab for
IHC
 control,
To: Tony Henwood <henwood@mail.one.net.au>,
HistoNet@Pathology.swmed.edu,
 Bruce A Rasmussen <brasmuss@osf1.gmu.edu>

Tony and Netters,
My sentiments exactly, that's one reason to always run a panel and not
single antibodies, this way we at least can derive some useful
information
from our negative controls.
Jeff SIlverman

----------
> From: Tony Henwood <henwood@mail.one.net.au>
> To: HistoNet@Pathology.swmed.edu; Bruce A Rasmussen
<brasmuss@osf1.gmu.edu>
> Subject: Re: omiting the primary Ab for IHC control,
> Date: Tuesday, March 02, 1999 5:48 PM
>
> It is a good idea to include a negative control with each case. Often
> when a panel of antibodies (eg S100, keratin and LCAg) are used the
> "negative" control is superfluous since one hopes that the tumour
> will be negative for one of the panel antibodies used.
> Tony Henwood
> www2.one.net.au/~henwood






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