I'll add a dissenting opinion here.  I have tried 5 different clones
of cyclin D1, including the Novocastra one (clone  P2D11F11), but my
favorite one is clone DR1BC2 (I purchase it from Biocare Medical, phone
800-799-9499, cat# CM-090D, but it may be available from other sources as
well.)  HOWEVER, I must add that ALL of the clones I tried (DR1BC2,
P2D11F11, DCS-6, 5D4, and AM29) do work, but that NONE of them is what I
would call a stellar performer, or an easy antibody to work with.  I
prefer the DR1BC2 clone because it gives me the most consistent
performance, and generally the strongest signal of any I have tried.
     For the rest, I do epitope retrieval in a steamer for 40 minutes in
pH 10 Tris, followed by a 20 minute cooldown.  I incubate in my primary
antibody @ 1:50 for 1 hour at room temperature, followed by secondary,
tertiary, DAB, etc. as per usual.
     And finally, I agree 100% with Zenobia Haffajee's final line, "Give
it a try, GOOD LUCK....wave your magic IHC wand before you start."

Phyllis Davie


>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!
>>
>>
>>
>>
>