We use Novo Castra's CD5 antibody Cat # NCL-CD5. We steam the slides for 20
minutes, cool for 20 minutes.  We incubate the primary for 1 hour at room
temperature followed by a 30 minute link, 30 minute label and 5 minutes in
DAB.  We were able to get a 1:25 titer.  Hope this helps

Joe Nocito, B.S., HT(ASCP)QIHC
Histology Supervisor
Christus Santa Rosa Hospitals
San Antonio, Texas


> -----Original Message-----
> From:	Dana Settembre [SMTP:settembr@UMDNJ.EDU]
> Sent:	Wednesday, December 01, 1999 8:17 AM
> To:	Trivett Melanie (PMC)
> Cc:	'histonet'
> Subject:	Re: CD5 immunohistochemistry
>
> On Wed, 1 Dec 1999, Trivett Melanie (PMC) wrote:
>
> > Dear Histonetters,
> >
> > I am keen to hear of success stories for immunohistochemistry of cyclin
> D1
> > and CD5 on formalin fixed sections.  Anyone willing to share protocols,
> > choice of primary antibodies?  Thanks in anticipation.
> >
> > Melanie.
> >
> >
> > Melanie Trivett, PhD
> > Department of Pathology
> > Peter MacCallum Cancer Institute,
> > East Melbourne, Victoria, Australia.
> >
> >
> >
> >
> >
> >
> Hello Melanie from Australia
>
> I use CD5 from NovoCastra Laboratories cat# NCL-CD5-4C7.  I pretreat with
> a citra buffer for 15 minutes in the microwave. I use the antibody at a
> dilution of 1:25 on my Dako autostainer along with Dako's LSAB2 kit. Other
> secondary and tertiary kits work fine too.  Your dilution may vary of
> course. I use tonsil for a positive control. Works nicely. Good Luck.
>
> Dana Settembre
> Immunohistochemistry Lab
> Pathology Department
> University Hospital
> Newark,  New Jersey
> USA
>