RE: p53 staining in cryosections
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From: | DORIT ZHARHARY <d_zharhary@sigma.co.il> (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
Reply-To: | |
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
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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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