Immunohistology on mouse using mouse monoclonals

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


In <Eyo5yD.9G6 at news.ki.se> nobody at nowhere writes: 
>
>Dear netter,
>
>To my dismay, the need has arisen to do some immunohistochemistry.
More 
>specifically I need to check out some muscle markers on our knock-out
mice. 
>The problem is that the antibodies for the markers are mouse
monoclonals, 
>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
got 
>similar staining using secondary antibody only as with any on the
antibodies. 
>
>I wonder what I might be doing wrong. It would seem logical to me that
the 
>background arises from endogenous mouse IgG that the secondary
antibody reacts 
>with. Could this be blocked somehow, maybe with unlabeled
anti-mouse-IgG? 
>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
aforementioned 
>literature gives only a very cursory description of the technique
used. 
>
>Sincerely,
>
>        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.

Tom





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