[Histonet] RE: Humanized mouse monoclonals

From:"C.M. van der Loos"

Dear Tom,
For the detection of a "humanized" monoclonal antibody on human tissue 
one need a procedure similar to the ARKit: in vitro labeling of the 
primary antibody with a biotinylated goat anti-IgG (of primary 
species) Fab-fragment. Such a kit is available from Molecular Probes 
and called a Zenon-kit 
(http://www.probes.com/products/zenon/zenon.html). According to their 
website it is available for mouse, rabbit and human primary antibodies 
and the label is either biotin or a fluorochrome.

Chris van der Loos, PhD
Dept. of Pathology
Academical Medical Center M2-230
NL-1105 AZ Amsterdam
The Netherlands

----- Original Message ----- 
>From  Thomas Crowell  
Date  Tue, 11 Jan 2005 12:41:40 -0500 
To  histonet@lists.utsouthwestern.edu 
Subject  [Histonet] Humanized mouse monoclonals 

Dear Histonetters,

I was wondering if anyone could provide me with a basic protocol using 
preformed complex of a humanized mouse monoclonal (human Fc ) with a 
biotinylated anti-human IgG to detect antigens in human cryostat 
- essentially, human on human IHC staining?  I have several antibodies 
with different total protein concentrations.


Tom Crowell
Cambridge, MA 

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