RE: mouse on mouse ELISA

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From:"Abizar Lakdawalla" <>
To:"'Connolly, Brett'" <>, "'HISTONET'" <>
Content-Type:text/plain; charset="iso-8859-1"

Hi Brett, the labeling (biotinylation) reagent and mouse block in our
iso-IHC kits may work well for mouse on mouse ELISA. We have had a similar
problem but with western blots, where the mouse antigen of interest had a
similar molecular weight to antibody light chain. On these western blots the
conventional procedure (using anti-mouse biotinylated secondary antibodies)
gave a very strong background band because the anti-mouse reacted with light
chain present in the tissue homogenate. We shifted to the iso-procedure
(mixed the mouse primary antibody with the labeling reagent followed by the
addition of the mouse block reagent - the whole mixture was then diluted and
added to the blot) resulting in the elimination of the non-specific bands.

Hope this helps

-----Original Message-----
From: Connolly, Brett []
Sent: Thursday, April 20, 2000 9:10 AM

A colleague is doing ELISAs with a murine monoclonal on murine peripheral
blood and is having background problems due to the secondary antibody. Are
than kits/reagents ala the mouse on mouse IHC kits that work in an ELISA
format? I'd be interested in hearing from vendors and any of you who have
done this.


Brett M. Connolly, Ph.D.
Merck Research Laboratories
Department of Pharmacology
PO Box 4
West Point, PA 19486
Ph. 215-652-2501
FAX 215-652-2075

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