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From:Debbie Pepperal <dpeppera@mail.newcastle.edu.au>
To:"Van Eyck, Deb" <deb.vaneyck@phci.org> (by way of Histonet)
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Hi Deb



We use the CyclinD1-GM from Novocastra, found it to be the best one so far.
CyclinD-1 is a tricky unstable antibody....likes to give problems. So donot
feel alone; going bald, well donot do it for the cyclind-1!
 Yes, sometimes it works great and at other times a bit iffy..
We microwave in an alkaline pH ( Trisbase at pH 10) for 20 mins on high in
a domestic microwave, allow to cool for 20 mins. Incubate with primary at a
1:25 dilution overnight at rt and use the ABC detection ; biotinylated goat
anti-mouse 1:200 for 30 mins and Streptavidin peroxidase at 1:500 for 30
mins at rt. DAB as the chromagen. Give it a try, GOOD LUCK....wave your
magic IHC wand before you start.


Zenobia Haffajee
HAPS, Newcastle, Australia


At 07:36 PM 11/04/00 -0500, you wrote:
>
>
>          Hi,
>
>               Would anyone who has Cyclin D-1 working reliably---and often
>please share all your info--including products used, times for all process,
>kit used etc-- and where is everyone finding mantle cell lymphoma for
>controls as that is what we are insterested in staining for????
>
>                     Thanks-Deb   I'm tearing my hair out on this one!
>
>
>
>



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