Re: cyclin D1

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From:amos brooks <>
To:Vinnie Della Speranza <>
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Hi Vinnie,
    We use Cyclin D1 frequently, on bone marrow biopsies. We use the DAKO antibody (M7155 ?) diluted to 1:50.
The antigen is best retrieved using a high pH antigen retrieval. (also from DAKO, I forget the pH) We use LSAB2
for a detection kit. Don't even bother with Envision+, it doesn't work. The incubation times following
peroxidase blocking are 20 min primary, 10 min Link2, 10 min Label2 then the chromogen you want. LSAB+ also
works but we phased this one out after a bad lot problem.
    The controls we use generally tend to be GI. We check each test we do to determine if it can be used as a
control. If it is positive with out ridiculous amounts of background then we cut extra slides to hold us over
until the next positive patient rolls along. If you are running low on controls, and a doctor asks for lymphoma
markers on a GI case and not Cyclin D1 run one anyway just to check, that is a good place to find them (Let the
doctor see it of course, never file a slide a doctor hasn't seen!)
    Just as a point of reference, I saw an incredible Cyclin D1 from a representative of SIGNET the other day.
(made me drool all over the microscope what a mess :-) I think theirs blew ours out of the water. There was
hardly any background staining, which is virtually impossible with this quirky antibody. Unfortunately I dont
know what their detection method was. :-(
good luck
Amos Brooks

Vinnie Della Speranza wrote:

> I would appreciate hearing from anyone who has this marker working reliably to learn
> 1. who's antibody you use
> 2. your antigen recovery method
> 3.  any caveats or tricks you might offer to provide better reliability
> 4. what you are using for a control
> we have had some cases stain very well, others that the pathologist insists should be positive that weren't.
> Vinnie Della Speranza
> Manager for Anatomic Pathology Services
> Medical University of South Carolina
> 165 Ashley Avenue
> Suite 309
> Charleston, SC  29425
> ph:  (843) 792-6353
> fax: (843) 792-8974
> email:

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