RE: [Histonet] alkaline phosphatase

From:"Bonnie Whitaker"

There are other alk phos chromogens available such as NBT/BCIP which yields
a blue-black color product, and Vector has several chromogens for HRP,
including a brick red, a purple, and it seems like some others.  I
personally like using the brick red HRP and the blue-black AP chromogens.
If you want to use 2 HRP protocols you can do as Kathleen suggested.
Bonnie Whitaker
-----Original Message-----
[] On Behalf Of Kathleen
Sent: Wednesday, February 18, 2004 1:54 PM
To: Martin Weber
Subject: Re: [Histonet] alkaline phosphatase

Yes, use plain DAB for your first AB and DAB nickel sulfate for your 
second. The procedure takes 4 days because you do them one at a time 
(with overnight incubation) You get brown for one and blue black for the 
other. Anyway, that is what we do.

Kathleen Spencer, HT
Lab Manager
On Wednesday, February 18, 2004, at 01:19  PM, Martin Weber wrote:

> Hello,
> I might need to do a dual staing procedure on paraformaldehyde fixed 
> fresh frozen sections. One marker will be stained with peroxidase 
> (DAB). As a second marker for IHC I read about alkaline phosphatase 
> (FastRed) which gives rise to a red color in addition to the brown 
> color of the peroxidase staining. To me this combination seems to be a 
> little bit unfortunate because the colors are sort of overlapping.
> Does anybody know of an alternative and complimentary method for dual 
> staining which might give rise to a different color other than red ? 
> Unfortunately due to special circumstances Immunofluorecence is not an 
> option.
> Any help is much appreciated. Thanks a lot !
> Martin Weber
> The Scripps Research Institute
> Dept. of Molecular and Experimental Medicine
> 10550 North Torey Pines Road
> La Jolla, 92037
> Phone: 858-784-8612
> Fax: 858-784-2030
> E-Mail:
> Martin Weber M.D.
> Work address:
> The Scripps Research Institute
> Department of Molecular and Experimental Medicine
> 10550 North Torrey Pines Road, Mail drop MEM 175
> La Jolla, CA. 92037, U.S.A.
> Phone at work: 001-858-784-8612
> Fax at work: 001-858-784-2030
> E-Mail:
> Home address:
> 7265 Charmant Drive, Apt 628
> San Diego, CA., 92122, U.S.A.
> Phone at home: 001-858-452-0544
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