[Histonet] RE:BrdU and GFP staining

[Pixley, Sarah (pixleysk)]

 Dear Steve:
I would stain for GFP with an antibody to GFP and do labeling with DAB.
I recommend the Elite ABC kits. The DAB is moderately resistant to the
BrdU penetration protocols. Then, fix your tissues well with
paraformaldehyde, i.e. 15 mins, 4%, and proceed with the BrdU, using a
system that labels with a fluorescent antibody. This will only work,
however, if your GFP is in different cells, or it is in the cytoplasm of
large cells that have no nuclear staining. The DAB can obscure the
secondary fluorescent labeling. 

Good luck. Stay Cool back at you.
Sarah Pixley,
Ohio

Message: 2
Date: Mon, 31 Jul 2006 10:22:41 -0700 (PDT)
From: Steven Coakley <sjchtascp <@t> yahoo.com>
Subject: [Histonet] BrdU and GFP expression
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <20060731172241.65699.qmail <@t> web38215.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Good afternoon from WI,
   
  We have several FFPE blocks from 2003 that express GFP.  Our lead
scientist would like to perform Brdu IHC in the same slides hoping to
express both.  As expected the AR removes the GFP green label.  Has
anyone any experience expressing Brdu and GFP on the same slide.
   
  Stay cool,
  
  Steve