[Histonet] Melanin and red chromogens for IHC

From:Gayle Callis

Vector have NovaRed, HRP substrate, it is a very dark brick red and
sensitive as DAB.   

AEC+ from DAKO or just their AEC is in ready to use form, with AEC+ being
quite sensitive, although not as much so as DAB and less so than Alk Phos
substrates other than NBT/BCIP (dark blue/purple).  The joy of AP
substrates is a pink red rather than orange red (AEC) or brick red (Vector
NovaRed).  You may want to try AP versus HRP for a tinctorial properties
your pathologist would like best. DAKO has a new AP chromogen that competes
with Vector Red, and this chromogen is permanent.     

Chris van der Loos has a wonderful chromogen chart in his book on Multiple
Staining methods, which gives some guidelines on primary antibody
concentration versus the sensitivity of chromogens.  We find it extremely
useful for both double staining and even single IHC staining. 

At 05:33 PM 4/7/2004 -0500, you wrote:
>There are also other chromogens available for HRP that aren't "DAB brown".
>I still like AEC, but Vector has several others now.
>Bonnie Whitaker
>-----Original Message-----
>From: histonet-bounces@lists.utsouthwestern.edu
>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
>Sent: Wednesday, April 07, 2004 4:24 PM
>To: LBlack@carilion.com; Histonet@lists.utsouthwestern.edu
>Subject: Re: [Histonet] Melanin Bleach & IHC
>instead of using the bleaching, or worrying that the dab looks like melanin,
>protect your tissue and the results by using alk phos fast red detection. it
>will stain red, is permanent and can be used on almost every antibody with
>the exception of ER/PR and if I recall correctly(ventana will tell me if its
>not) Alk Phos fast Red is a bit more sensitive for detection than dab, the
>pathologist will adjust i assure you and morphology is saved.          dana
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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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