[Histonet] Counterstain for dual chromogenic

[Rene J Buesa]

You could do that or you could use DAB for one antibody ("brownish" color), followed by DAB + Nickel for the other ("bluish" color) and they are both permanent and alcohol resistant.
René J.

--- On Tue, 4/12/11, Eva Permaul <eca9 <@t> georgetown.edu> wrote:


From: Eva Permaul <eca9 <@t> georgetown.edu>
Subject: [Histonet] Counterstain for dual chromogenic
To: histonet <@t> lists.utsouthwestern.edu
Date: Tuesday, April 12, 2011, 11:37 AM


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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