[Histonet] RE: human Ab on human tumor

[C.M. van der Loos]

Dear Wen,
I think your observation is completely right. When doing human-on-human staining the endogenous immunoglobulins in the tissue section will cause the background. In your case there will be no human immunoglobulins around and indeed this yields a background-free result. If you want to absolutely sure your result isn't fake, you may label your primary antibody directly. Most convenient is a Zenon kit (http://www.probes.com/products/zenon/zenon.html). The labeling procedure with biotin- or Alexa fluorochromes labeled Fab fragments is comparable with the Dako ARKit for mouse-on-mouse staining. 

Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center
Amsterdam - The Netherlands

----- Original Message ----- 
>From  wen eng <weneng2004 <@t> yahoo.com> 
Date  Tue, 26 Oct 2004 12:00:56 -0700 (PDT) 
To  histonet <histonet <@t> lists.utsouthwestern.edu> 
Subject  [Histonet] human Ab on human tumor 
Hi histonetters,
 
Currently I am doing IHC using human Ab on human tumor. I know it would cause "messy" background. But this tumor was developed by injection human tumor cell to nude mice. The results came out pretty good. I didn't use conjugated primary Ab, but just traditional method. I think it's because the tumor was developed in mice, the tissue would not contain endogenous human serum that causes the "messy background", although the tumor is human tumor. Am I right on this? Or my stain result just "fake"?
Hope I said it clearly. Could any expert give me help?
 
Thanks in advance!