If you have not already done a literature search I would recommend starting
there to see if anyone has a protocol already developed. If not I like to
start with the intended target to make sure that the antibody works as
intended. An antibody on a human protein against the recommend tissue and
fixation. Following that I like to try the animal tissue fresh, fresh formal
fixed and formalin fixed paraffin embedded[FFPE]. Sometimes you get
incredibly lucky and it work on FFPE tissue of the species you are
interested and it seems very simple. More frequently it requires lots on
innovative tweaking and possibly and entire bust!! Depends on how much
effort you want to but into developing the protocol.

Good luck - I hope it is easy!

Cynthia Favara
NIAID/NIH/RML/LPVD
903 South 4th Street
Hamilton, MT 59840
406-363-9317


-----Original Message-----
From: sanjeev francis [mailto:sanjeevfrancis@hotmail.com]
Sent: Thursday, July 31, 2003 12:00 PM
To: histonet@pathology.swmed.edu
Subject: Immunostaining of murine aortic cross-sections


I am interested in immunostaining mouse aortic cross-sections to look for 
the tissue distribution of a particular protein.  I will be using a 
polyclonal Ab.  My question is how to prepare the tissue sections.  I have 
read that frozen sections preserve the antigenic integrity of tissues but 
have also heard that paraffin fixation is "milder" on the tissue.  Which 
technique should I use to optimize my immunostaining?

Thanks,
Sanjeev Francis, MD
Research Fellow
Weill Medical College of Cornell University

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