[Histonet] CD26

[Orr, Rebecca]

Has anyone ever stained for CD26 in FFPE human tissue?  If so, please
share your experiences.  I'm supposed to get this stain up and running
for a clinical trial that is starting up and I'm not having much luck.
The Japanese group we're working with is coming to town in a few weeks
and I'm getting a little nervous. 

I need to stain in kidney and some other normal tissues, but I want to
see it in tonsil first so I can really have some faith in the stain.  I
have several antibodies, including the humanized version that is the
drug.  The humanized antibody is biotinylated.

Thanks,

Mark Adam Tarango HT(ASCP)
Histology & IHC Supervisor
Nevada Cancer Institute
One Breakthrough Way
Las Vegas, NV  89135
Direct Line (702) 822-5112
Treo (702) 759-9229
Fax (702) 939-7663
  

Mark,
I just started working up CD26 this week, using the rabbit polyclonal
catalog # sc9153 from Santa Cruz.  The brief data in the 2006 catalog
indicates this would have been   the most appropriate antibody to use on
FFPE tonsil.
However, after I purchased the product, I went onto their website
www.scbt.com  and found that the description of this particular rabbit
ab had changed and it was no longer recommended for FFPE.  They did have
a mouse ab that was described as working with FFPE.  In hindsight, I
would have most likely purchased the mouse ab first off with this
information.  

My result from this rabbit ab is "decent" the stain is not as well
defined as I'm used to seeing on CD T cell markers, I may ask them for a
sample of the mouse just to see if that's the problem.

I have results with a dilution range of 1:200 to 1:500 on tonsil
(depends on how strong you'd like the staining) Dilute with pbs based
diluent (biocare green) HIER was done with Reveal and Borg, (biocare) in
a decloaker The Borg (hi pH) gave a more distinct stain but the reveal
had better morphology....
Detection was Mach4/DAB (biocare)

Hope this helps.
Becky

Becky Orr CLA,HT(ASCP)QIHC
Anatomic Pathology
Evanston Northwestern Healthcare
847-570-2771