Re: immunodetection of peroxidase in brain sections

From:"J. A. Kiernan" <jkiernan@uwo.ca>

On Thu, 12 Apr 2001 sthomaska@yahoo.com 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
   kiernan@uwo.ca
   http://publish.uwo.ca/~jkiernan
 




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