Re: [Histonet] re: Background on FFPE mouse tissue

From:Gayle Callis

Good point, rat antimouse, hamster antimouse, or rabbit antimouse (if they 
have a monoclonal for mouse made in rabbit) or rabbit antihuman.

Not only secondaries adsorbed to mouse, but also use mouse serum in 
diluents/normal serum blocking. We prefer F(ab')2 frag of IgG secondaries, 
adsorbed to mouse in order to eliminate fc receptor binding.  Jackson has 
excellent antibodies for that purpose.
  Many of the kits for clinical use contain multilinks for mouse, rabbit 
and goat.  If you use a single link polymer kit for rodents, goat, rabbit, 
you will eliminate biotin and the mouse on mouse problems.  Biocare Promark 
single links work very well since they have an array of single links useful 
to research.  I believer Vision Biosystems also has this available.

If it is a mouse on mouse problem and you must use a mouse monoclonal, then 
kits that biotinylate the primary are superb.  DAKO ARK and Biocare have 
excellent kits for this purpose.  A lady gave an excellent poster on this 
at NSH a few years back, and her results were very convincing for the use 
of this kind of kit, no secondary antibodies are used at all.

At 11:17 AM 9/17/2007, you wrote:
>As Paul suggested, my first thought is that the background is due to the
>use of mouse monoclonals. A mouse-on-mouse kit may help, but my limited
>experience with these has not been very positive; they may be a bit
>unpredictable, and/or cause a marked decrease in the signal. Much
>preferable is to use primary antibodies from another species.
>Jean-Martin Lapointe, DMV, MS, dACVP
>AccelLAB Inc
>Histonet mailing list

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610

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