[Histonet] RE: Can Alk phs be run on peroxidase slides?
Basically I think this is possible. In the end it's a kind of
sequential double staining. Some remarks:
* Of course you have to be aware of animal species: if your AP stain
is another mouse primary you may end up with a cross-reactions.
This type of cross-reaction is not really expected since DAB was
used as chromogen. DAB reaction product has the unique
characteristic of effectively 'sheltering' all immunoreagents used
in the first staining sequence.
* If you want to be on the safe side with cross-reactions, perform
HIER (using the same buffer as for the first one) for just 5 min +
10 min cooldown. This effectively removes or at least destroy all
your first step reagents while the DAB reaction product remains
* A second streptavidin-biotin detection system is not recommended.
To be on the safe side use a AP-labeled polymer. In case you
really need another streptavidin-biotin detection system, perform
a biotin blocking first (avidin 0.1%, 15 min - d-biotin 0.01%, 15
min, or a commercial biotin blocking kit).
* Given for example that your first DAB stain is cytoplasmic and the
second AP stain is nuclear, AP in red with Liquid Permanent Red
(Dako) is a good choice.
* Destaining of a DAB-stained slide will be a very hard job, not to
Lots of success,
Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
NL-1105 AZ Amsterdam
Date: Mon, 24 Jul 2006 12:44:48 -0400
From: "Dana Settembre"
Subject: [Histonet] Can Alk phs be run on peroxidase slides?
One of my pathologists is asking if I can take a slide that has
been stained with a
peroxidase/avidin/biotin/DAB detection system and re-stain with an
Alkaline Phosphotase detection system.
Can it work?
Must it be de-stained?
Any info will help. Thanks,
University Hospital - UMDNJ
Histonet mailing list
<< Previous Message | Next Message >>