RE: Immunos on Mouse tissue
I am curious, I have been unsuccessful with CD31 from Pharmigen. Trying to
use it for MVD study on mouse brain fixed in paraformaldehyde. Do you HIER.
Do you digest the tissue (I think recommended)? What dilution do you use? I
presume that this is the rat primary. Do you really need Tyramide Signal
From: Barbara Wright [mailto:bwright@gene.COM]
Sent: Thursday, January 31, 2002 12:40 PM
Subject: Re: Immunos on Mouse tissue
If you are using the DAKO CD31 antibody - it will not react with mouse
endothelium. Pharmigen CD31 (MEC13.3) works well on FFPE with Vectastain
ABC followed by NEN TSA-kit. And our Collagen IV is from a polyclonal from
Chemicon. Both work well on FFPE mouse tissues.
> I am in need of some help? I am trying to do immunos on mouse tissue. I
> currently use the Dako LSAB+ kit and I know that some of my antibodies are
> monoclonal. I have tried the Dako ARK kit and have not had any luck. I
> know what I may have done wrong. I also have tried the Dako Envision
> polyclonal kit. I have changed my antibodies from mono to poly or goat. I
> have tried different retrieval methods. What can I do now? Any help would
> appreciated. The two antibodies I am working with mostly is the Cd31 (goat
> polyclonal and Rat anti mouse) and Collagen 4 (monoclonal and polyclonal).
> Donella Stillings
> University of Colorado HSC
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