Hi Ron,

We've recently switched to Biogenex' monoclonal CD3 and like it a lot.  We
use it on Ventana immunostainers but have also seen it demo'd on the
Optimax.  Our pretreatment regime is HIER:  20" @ 92-95 degrees C in a water
bath in 1mM EDTA, pH 8 or 2.5" in the same buffer in the Biocare Decloaking
Chamber.  We use a dilution of 1:10 and an incubation time of 32" at 42
degrees C followed by Ventana Amplification (rabbit x mouse IgG, mouse x
rabbit IgG).  Compared to most of our other antibodies, this one  takes
quite a bit thrown at it to get it to optimally stain but the results are
excellant.

Hope this helps; sorry we use a different vendor's Ki67.

Linda A. Sebree, HT
University of Wisconsin Hospital & Clinics
Immunohistochemistry/In Situ Hybridization Laboratory
D4/218-2472
600 Highland Avenue
Madison, WI  53792-2472


(608)265-6596
FAX: (608)263-1568

> -----Original Message-----
> From:	Pathrm35@cs.com [SMTP:Pathrm35@cs.com]
> Sent:	Friday, February 25, 2000 2:13 PM
> To:	Histonet@pathology.swmed.edu
> Subject:	ki-67 and cd3 immunos
> 
> Fellow Histonetters,
>    I am having problems with CD3 and Ki-67 immunos staining correctly. I
> am 
> using 
> Biogenex antibodies and have tried their procedure by hand and on their 
> immunostainer.
>    I have had mixed results with the Ki-67 and no results with the CD3,
> both 
> on the stainer and by hand. If anyone has any suggestions I greatly 
> appreciate it.
> 
>    Ron Martin, B.Sc. HTL,HT (ASCP)
>    Palm Beach Pathology
>    West Palm Beach, Fl