RE: Desmin on BM BXs

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From:"Hewlett Bryan (CMH)" <HEWLETT@EXCHANGE1.CMH.ON.CA> (by way of histonet)
To:histonet <>
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In my experience anti-Desmin, clone D33, works well on B5 fixed tissue
and on zinc formalin fixed/decalcified tissue. It does not respond well
to proteolytic digestion( ficin, bromelain, trypsin,pronase or pepsin),
nor does it particularly like HIER, with any of the six buffers that we
have tested. On NBF fixed tissue, after proteolysis, staining was almost
completely abolished, with the exception of bromelain  After HIER,
staining was diminished in all cases. We use D33 from Dako, with NO
pre-treatment, diluted 1:50 for 1hour at R.T., followed by LSAB.

Hope this helps,



>From: 	Sebree Linda A.[]
>Sent: 	December 18, 1998 10:23 AM
>To: 	'Histonet'
>Subject: 	Desmin on BM BXs
>Hi Histonetters and Happy Holidays,
>We're having problems getting desmin to work on B5 fixed BM BXs.  We've been
>using a bx that is full of tumor cells metastasized from a strongly desmin
>positive primary.  We routinely use Neomarkers antibody, clone D33, at a
>1:30 dilultion after 20" EDTA HIER on formalin fixed tissue.  So far,
>everything I've tried (combinations of heat retrieval and enzyme treatment
>as well as each alone) has resulted in negative results.  Any help out
>Linda Sebree, HT
>University of Wisconsin Hospital & Clinics
>Department of Laboratory Medicine
>IHC/ISH Laboratory
>600 Highland Ave.
>Madison, WI  53792-2472
>Phone:  (608)265-6596
>FAX:  (608)263-1568

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