From:Tony Henwood

If these are air-dried, acetone treated smears, why are you retrieving them? No formalin fixation is being used.

Tony Henwood JP, BappSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital at  Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: (02) 9845 3306
Fax: (02) 9845 3318

-----Original Message-----
From: []
Sent: Friday, 1 November 2002 4:25
Subject: TRAcP

Hello fellow Histonetters:

       We have started experimenting with Zymed's TRAcP antibody with poor results on peripherial smears.  Does anyone have any guidelines to follow to bring this antibody up and running?  Washing off is a problem as well.

1.  Fix in Acetone for 1 minute.
2.  Steam retrieval in EDTA for 30 minutes.
3.  Cool for 30 minutes.
4.  Primary antibody for 60 minutes.
5.  Link for 30 minutes  (LSAB2)
6.  Strep for 30 minutes.
7.  DAB for 5 minutes
8.  Counterstain with Hema for 5 - 8 seconds.

The procedure is run on DAKO Autostainer.

       Thanks everyone for your help.

Jim Bradford
Orlando, FL

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