RE: p53 staining in cryosections

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From:DORIT ZHARHARY <> (by way of histonet)
To:histonet <>
Content-Type:text/plain; charset="iso-8859-1"

The fixation method recommended for frozen sections using this antibody is:
1. Methanol / Acetone (1:1 v/v), pre cooled at -20oC, 10 minutes, followed
by PBS wash.
2. Try prolonging the incubation time with the antibody, even overnight at
4oC, and try several antibody dilutions.

Dorit Zharhary

From:  Pat Eden
Sent:  יום רביעי 03 מרץ 1999 18:55
To:  'HistoNet Server'
Subject:  p53 staining in cryosections

I am looking for tips on immunohistochemical staining of p53 in frozen
sections of colon tumors.  I am working with an antibody (Sigma
BP53.12)which works well with paraffin sections, however, it doesn't seem
to be as effective with frozen sections.  I am fixing the tissue in cold
acetone for ten minutes, and using a Sigma ExtrAvidin peroxidase DAB
detection system.  Any advice on immunohistochemical protocols on frozen
sections or p53 antibodies would be greatly appreciated.

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