Hi all, Biocare also suggests using 4 micron sections or thinner.
Marianne




On Thu, 18 Feb 1999, Phyllis Davie wrote:

> Hoo-Boy!  Wouldn't I love to hear that information too.  I have just
> placed an order for my 5th clone of Cyclin D1.  They all "work"
> (sort-of), but none at a level that I would call good.  I'll give you a
> run-down of what has worked for me, but I would sure like to hear from
> others too.
>
> clone P2D11F11 (Novocastra cat# NCL-CYCLINC1-GM) 1:250.
>  Pretreat by steaming 20 minutes in 6mM citrate, pH 6.0, followed by a 20
> minute cooldown.  Rinse in water, then digest 10 minutes in 0.1% pronase
> (w/v in PBS) at room temperature.  Rinse again, then proceed.  This
> treatment hereafter referred to as S20/+10.
>
> clone DCS-6  (Dako cat# M7155)  1:100  S20/+10
>
> clone 5D4   (Immunotech-Coulter cat# 1740) 1:1000  S20/+10
>
> clone DCS-6 (same clone as DAKO, but from NeoMarkers, cat# MS-210-P0)
> 1:500, S20/+10
>
> clone AM29   (Zymed cat# AS18-0220)  1:100,  Pretreatment in a microwave
> pressure cooker in the citrate for 10 minutes at pressure, followed by
> cooldown until 60 degrees C.
>
> clone DR1BC2   (	BioCare Medical	CM090A or CM090D)  ON ORDER (the
>company
> recommends using their "EDTA Decloaker" antigen retrieval buffer for best
> results)
>
>
>    I have tried a wide variety of other pretreatment options with varying
> buffers, pH's, times, heat, no-heat, enzymatic digestions, ... anything I
> could think of.  The most recent thing I heard was that a Tris/EDTA
> buffer at pH 8.5 would do the trick, but I don't know concentrations of
> the salts.  We gave it a try with 100mM Tris and 10mM EDTA for 20 minutes
> in the steamer.  It gave results that were "working", but no better than
> my S20/+10.  If anyone knows the magic antibody or pretreatment, I would
> dearly love to hear about it.
>
>
>
>
>
>
>
> >Anyone have any recommendations for good results with cyclin D1. Which
> >company's antibodies and which method works best for you?
> >Thanks in advance, J:>)
> >JOYCE WEEMS
> >PATHOLOGY LAB MANAGER
> >
>
>
> Phyllis Davie
> Clinical Laboratory Supervisor
> PhenoPath Laboratories
> Seattle, WA
> pdavie@phenopath.com
> (206)374-9009  phone
> (206)374-9009  fax
>
>
>
>