[Histonet] bone tissue falling off after AR step

[patsy ruegg]

Brett,
Bone is hard and the cortical bone will be the most difficult to preserve.
For IHC on bone I airdry the sections overnight before baking at 58dc for at
least 1 hr.  I try to avoid HIER and use enzyme digestion if possible for
antigen retrieval.  If I do need to use HIER, I use a waterbath no higher
than 72dc for 3 hrs. or 32dc overnight.
Patsy

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Connolly,
Brett M
Sent: Tuesday, December 19, 2006 7:57 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] bone tissue falling off after AR step

Hey boneheads,
A colleague is having problems with cortical bone detaching from the
slide during or after the antigen retrieval step. He is working with
mouse tibia. 

He uses Superfrost+ slides. The AR is with citrate buffer (pH6),
performed in a waterbath for 10 minutes. He bakes the slides for 15 min
at 56C prior to deparaffinizing.

What do you suggest?

Thanks, Brett

Brett M. Connolly, Ph.D.
Research Fellow
MRL, Imaging Research
Merck & Co., Inc.
WP-44K
PO Box 4
West Point, PA 19486
PH 215-652-2501
fax. 215-993-6803
e-mail. brett_connolly <@t> merck.com


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