Immunohistology on mouse using mouse monoclonals

Thomas R. Anderson babco at
Sat Sep 5 11:06:13 EST 1998

In <Eyo5yD.9G6 at> nobody at nowhere writes: 
>Dear netter,
>To my dismay, the need has arisen to do some immunohistochemistry.
>specifically I need to check out some muscle markers on our knock-out
>The problem is that the antibodies for the markers are mouse
>which makes their use on mouse tissues dubious at least to my naive 
>understanding. Since these particular antibodies were used
successfully on 
>mice recent literature I decided to give it a try. Unfortunately, I
>similar staining using secondary antibody only as with any on the
>I wonder what I might be doing wrong. It would seem logical to me that
>background arises from endogenous mouse IgG that the secondary
antibody reacts 
>with. Could this be blocked somehow, maybe with unlabeled
>Where would such reagent be available?
>If you could point me to some direction, I'd be most grateful. For
instance, a 
>reference to a good technical review would be most useful, the
>literature gives only a very cursory description of the technique
>        Jarkko Kortesmaa
>        Karolinska Institute
>        Stockholm, Sweden
>        (To reply by email, remove nospam from adress.)        
Hello Jarkko--

I think the trick here is to directly label the primary antibody (with
HRP or biotin, for example) thereby avoiding the use of the secondary
antibody.  Use of an anti-mouse secondary reagent in a mouse system
will give you loads of background, as you have found.

Good luck.


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