[Histonet] Making DAB+ in Dako Ark darker

From:Gayle Callis

You can make the brown endproduct a very dark, black brown (almost black!)
by using the DAKO DAB enhancer.  This works beautifully with their DAB+
that is in ARK kit or you can always do another enhancer to turn the
product black.  This has been discussed at great lengths in Histonet
archives.  Other companies have enhancers ready to use, Zymed and Vector,
but I am not sure if they are black.   

Since the kit is a kit, we prefer to use the DAKO enhancing product, pretty
much optimized for their DAB+ which (with their enhancer), allowed us to
get a murine CD4 diluted out 1:15,000 (0.5mg/ml)and more.  This was not an
ARK kit method, but we were impressed with the overall chromogen/enhancer.
We had to be careful about any residual peroxidase or pseudoperoxidases in
tissues when using DAKO DAB enhancer.  We used the glucose oxidase
endogenous peroxidase method for quenching with very clean results.

At 01:25 PM 9/25/2003 +0100, you wrote:
>          Although the brown reaction product with Dako Ark looks lovely
>I'd like a black product. Has anyone used intensifying techniques with Dako
>Ark and can they offer any hints and tips.
> Ian.
>  Dr. Ian Montgomery,
> Histotechnology,
> Graham Kerr Building,
>& Life Sciences,
> University of Glasgow,
> Glasgow,
> G12 8QQ.
> Tel: 0141 339 8855 
> Office: 4652
> Lab: 6644.
> Pager: 07625 702883
> e-mail: ian.montgomery@bio.gla.ac.uk  
Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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