Re: immunodetection of peroxidase in brain sections

From:"J. A. Kiernan" <>

On Thu, 12 Apr 2001 wrote:

> I need to double label a tracer (WGA-HRP) and a cell
> specific marker. I assume in this case WGA-HRP
> detection cannot be done using peroxidase reaction. 

You could detect the lectin-HRP conjugate by virtue of
its peroxidase activity, using DAB and H2O2 in the
usual way, and then detect the cell-specific antigen 
with one of the methods that uses alkaline phosphatase 
rather than peroxidase as the labelling enzyme. 
Alkaline phoshatase methods typically give purple 
deposits; a colour that contrasts well with the brown
of oxidized DAB. Usually you have to use a water-miscible 
mounting medium.
John A. Kiernan
Department of Anatomy & Cell Biology
The University of Western Ontario
London,  Canada   N6A 5C1

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