Richard,

CD10(56C6) is extremely robust but, as you correctly point out, fixation
sensitive.
We use it at 1:80 for 1 hour @RT on tissue fixed in Zinc formaldehyde
and de-mineralized, also on B5 fixed material. However, for tissue fixed
in NBF we find it necessary to incubate overnight @RT, at a dilution of
1:40. We did our initial titreing trials on both small bowel(brush
border is positive) and on kidney( glomeruli and prox tubules) fixed in
NBF, ZF and B5 all with and without de-mineralization, in order to
arrive at the dilution and time which gave matched intensities
regardless of fixation.

Regards

Bryan

>----------
>From: 	Richard Cartun[SMTP:Rcartun@harthosp.org]
>Sent: 	August 9, 1999 12:41 PM
>To: 	Histonet@pathology.swmed.edu
>Subject: 	CD10
>
>Like some of you we have been having problems with getting CD10 (clone 56C6)
>to work on formalin-fixed tissue.  We tried incubating for 30 minutes at RT,
>but found that we had to use a 1:10 - 1:20 dilution in order to get any
>immunoreactivity whatsoever. 
> At those concentrations the antibody is too expensive to use.  Therefore, we
>have been incubating overnight at a dilution of 1:100.  Last Friday I
>evaluated a consult case that showed 4+ (maybe even 5+!) CD10
>immunoreactivity.  Why did this case stain so
> strong?  The tissue was fixed in B5.  Our hematopathology fellow brought to
>my attention an article that was published in 1998 where the authors found
>that CD10 worked better in B5-fixed tissue (see Mod Pathol
>1998;11(11):1046-1051).  I don't know if all
> B5-fixed tissue will stain this nicely, but it sure was gratifying to see
>"dark" CD10 immunoreactivity for once.
>
>RWC
>
>