RE: Cyclin D1 antibody

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From:"Nocito, Joseph" <joseph_nocito@srhc.iwhs.org>
To:"'Brennan, Liam'" <Liam.Brennan@bll.n-i.nhs.uk>, HISTONET <histonet@pathology.swmed.edu>
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Liam,
we have been using Cyclin D-1 from Biocare Medical, Walnut Creek,
California, USA, telephone number 1-800-799-9499 with their EDTA HIER
solution and have been getting good results.  We steam the slides in EDTA
for 20 minutes, cool for 20 minutes.  We perform a 30 minute incubation in
the primary antibody, linking antibody and the label, with a 5 minute
incubation in DAB.  Hope this helps

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


> -----Original Message-----
> From:	Brennan, Liam [SMTP:Liam.Brennan@bll.n-i.nhs.uk]
> Sent:	Tuesday, June 27, 2000 9:12 AM
> To:	HISTONET
> Subject:	Cyclin D1 antibody
> 
> 
> 	Histonetters-   Has anyone out there got a reliable, method for
> the demonstration of Cyclin D1 in human tissues. We currently use the
> DAKO  monoclonal Cyclin D1 antibody raised in mouse, following heat
> mediated antigen retrieval in citrate buffer pH 6.0 and we are currently
> getting variable results. We have also tried a high pH(9.8-9.9)
> retrieval solution from DAKO, but with similar results.  All
> contributions welcome.
> 
> 			Liam Brennan
> 			Histopathology Department
> 			Belfast City Hospital
> 			Northern Ireland



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