[Histonet] RE: [IHCRG] ALK-PHOS> staining of OCT?

[Connie McManus]

You don't need to use DAB... you can use AEC.  It has a nice dark red
stain. This is soluble in ETOH and xylene, so you need to use an aqueous
mounting media.  

Connie McManus
Utah Veterinary Diagnostics Laboratory
Utah State University
Logan, UT
Phone:  435/797-1891
fax: 435/797-2805


-----Original Message-----
From: rfail <@t> toolkitmail.com [mailto:rfail <@t> toolkitmail.com] 
Sent: Monday, May 24, 2004 9:55 AM
To: Connie McManus
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [IHCRG] ALK-PHOS> staining of OCT?

 Mel-5 is for melanoma and stains melanocytes so DAB is not
an option


 Have you tried using HRP instead of AP?  I had a similar
> problem with gut tissue on BVD (Bovine Viral Diarrhea)
> except that in addition to this blush, I also got this
> gorgeous red outline of all the microvilli even after
> levamasole quenching... not what you want to see, but it's
> artistically appealing.  I concluded that gut is not a
> good tissue to use with AP.  There are some tissues that
> have too much endogenous AP and quenching (blocking) just
> isn't enough... you need to use a different detection
> system.  I don't really know if this is your problem or
> not, but it's one thing you might want to consider.
>
> Connie McManus
> Utah Veterinary Diagnostics Laboratory
> Utah State University
> Logan, UT
> Phone:  435/797-1891
> fax: 435/797-2805
>
>
> -----Original Message-----
> From: Rena Fail [mailto:rfail <@t> charleston.net]
> Sent: Monday, May 24, 2004 2:37 AM
> To: histonet <@t> lists.utsouthwetern.edu; Ihcrg
> Subject: [IHCRG] ALK-PHOS> staining of OCT?
>
> Frozen sections stained with Mel-5 using an alkaline
> phosphatase procedure are showing a pink blush surrounding
> the tissue on the slide.I washed the slides in running
> water for 30 minutes, then rinsed in buffer (TRIS),
> applied all our usual blocks, protein block, biotin
> blocking reagents, and levamisole, it's still there. The
> intensity is exactly the same in the lower dilutions as in
> the higher. It doesn't interfere with the interpretation
> of the stain, but our docs are used to very 'clean" IHC
> slides for DAB and ALK-Phos. I used treated slides. Any
> advice on how to get rid of this blush would be
> appreciated.
>
> Rena Fail, HT(ASCP)QIHC
>  IHC/SS
> Medical University of SC
> 165 Ashley Ave.
> Charleston, SC 29425
>
>
>
> [Non-text portions of this message have been removed]
>
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