Re: p21

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From:Kappeler Andreas <> (by way of histonet)
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Hi Linda

No experience with Santa Cruz' p21, nor with Ventana (staining by hand; we
are old fashioned, I know). However reasonably good results with 2
monoclonal p21 antibodies, i.e.
Clone DCS-60.2, NeoMarkers, # MS-230-P; working conc. 4.0 ug Ig/ml;
pretreatment in 10 mM citrate buffer pH 6.0, pressure cooker with 3 min. at
full pressure. Visualization is with a StrABC/AP system.
Clone EA10, Oncogene, # OP64; working conc. 4.0 ug Ig/ml; pretreatment in 50
mM Tris with 5% Urea, pH 9.5, microwave oven; visualization as above.
Good luck,

Andi Kappeler, Ph.D.
Institute of Pathology, University of Bern, Switzerland

Original message:
> Hi 'netters,
> Does anyone out there have a protocol for p21 (also designated WAF1/Cip1)
> for use on FFPE tissue; preferably on a Ventana automated immunostainer?
> I have Santa Cruz's antibody (cat. #sc-8170) and they (SC) recommended
> with citrate buffer, 1:50 to 1:100 dilution and colon CA as a positive
> control.  My first attempt yielded negative results and as this is a
> research project, all the optimization time and cost is borne by the
> researcher and I'm trying to keep his costs down by not reinventing the
> wheel.
> Thanks in advance,
> Linda A. Sebree, HT
> University of Wisconsin Hospital & Clinics
> Immunohistochemistry/In Situ Hybridization Laboratory
> D4/218-2472
> 600 Highland Avenue
> Madison, WI  53792-2472
> (608)265-6596
> FAX: (608)263-1568

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