[Histonet] Re: Histonet Digest, Vol 89, Issue 12

[Mark Elliott]

Date: Tue, 12 Apr 2011 11:37:17 -0400
From: Eva Permaul <eca9 <@t> georgetown.edu>
Subject: [Histonet] Counterstain for dual chromogenic
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <4DA471AD.3040002 <@t> georgetown.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

Eva
 
I have been doing some double labelling using Vulcan Fast Red from Biocare and  Vector Blue Alkaline Phosphatase Substrate Kit III from Vector Labs.  For counter stain I use Vector Methyl Green and the combination works great and looks quite nice.
 
Mark
 
Good morning,
I have been attempting to do some dual chromogenic staining. I am using 
a mouse and a rabbit antibody. For the mouse antibody I am using an 
HRP-mouse secondary followed by AEC for visualization. For the rabbit 
antibody I am using a biotinylated rabbit secondary, followed by ABC-AP 
and vector blue. My problem is what to use as a counterstain. From 
everything I have read so far there isn't one that would work without 
having some problems. I was thinking of trying a very light Hematoxylin 
and see if it doesn't disrupt the vector blue visualization too much. 
Does anyone else have any suggestions?
Thanks,
Eva

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