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@charleston.net]
> Sent: Monday, May 24, 2004 2:37 AM
> To: histonet@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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