RE: CD10 ???

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From:Hewlett Bryan <HEWLETT@HHSC.CA>
To:histonet <>, "'Amos Brooks'" <>

Amos and Marylou,

After extensive testing, we settled on Novocastra CD10, clone
56C6(NCL-CD10-270), 1:80 following HIER @ 98C in citrate buffer pH6.0 for 20
min. For FFPE sections, incubation is overnight @RT, for B5 or AZF fixed
sections, 1 hour @ RT. Detection is by LSAB, chromogen AEC. With detection
by EnVision, dilution can be pushed to 1:160. This antibody works extremely
well for us, even after demineralization.
We performed our initial titrations on sections of normal kidney, small
bowel, tonsil and bone marrow, with the fixatives above, both with and
without demineralization.  We found very little immunoreactivity in normal
tonsil and marrow, what there was being of low intensity. A full range of
intensity was seen in kidney and bowel, we settled on kidney as a procedural
control for CD10. Further validation studies on neoplastic lymphoid lesions,
confirmed our dilutions etc.  We do see very intense reactions in many ALL
cases, we consider this to be the optimum. We have seen no false positives,
as confirmed by flow cytometry, occasional false negs, usually on specimens
that have been rushed through, but that's NO surprise!!!

For CD5, Novocastra, clone 4C7, for FFPE sections dilution 1:25 following
HIER @ 98C in EDTA pH 8.0. Incubation is 1hour @RT, detection is either LSAB
or EnVision. 

Best Regards,


> ----------
> From: 	Amos Brooks[]
> Sent: 	April 29, 2000 2:46 PM
> To: 	histonet
> Subject: 	CD10 ???
> Hey All,
>     One of our hematopathologists asked for an an antibody to be worked
> up that we dont have yet. Does anyone have any suggestions for CD10 on
> formalin fixed paraffin embedded sections. Please include which
> detection kit you use and any pretreatments and titrations you use.
> Thanks much,
> Amos Brooks

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