RE: [Histonet] Decal Bone Antigen retrieval

From:"patsy ruegg"

I avoid hier and especially in a pressure cooker for bone IHC.  Like Liz I
prefer enzyme digestion to hier for bone, if I have too, I use a gentle
steamer or waterbath and not a pressure cooker.

Patsy Ruegg, HT(ASCP)QIHC
12635 Montview Blvd. #216
Aurora, CO 80010
fax 720-859-4110 

-----Original Message-----
[] On Behalf Of Liz Chlipala
Sent: Tuesday, January 22, 2008 12:18 PM
To: Igor Deyneko;
Subject: RE: [Histonet] Decal Bone Antigen retrieval


Its going to depend upon what antibody you are trying to detect but if
possible I would try enzyme retreival over HIER, my favorite is
proteinase K, but we also use pepsin, pronase, chondroitinase, etc.  It
depends upon the antibody.  If you have to use HIER you can its just
thats it likely you are going to loose portions of the tissue.


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, CO 80308
phone (303) 735-5001
fax (303) 735-3540
Ship to Address:
Premier Laboratory, LLC
University of Colorado at Boulder
MCDB, Room A3B40
Boulder, CO 80309

-----Original Message-----
[] On Behalf Of Igor
Sent: Tuesday, January 22, 2008 12:01 PM
Subject: [Histonet] Decal Bone Antigen retrieval

Dear histonetters!
I have a question. Recently I have received decaled mouse hind paw joint
slides, which I have to do IHC on. Can anyone advise an antigen
retrieval solution and a method. The current method I use and has been
working fine in the past is as follows:
Either Citrate or DAKO Retrieval solution in a conventional pressure
cooker for 20 minutes , then I would let it stand with the open lid for
another 10 minutes, and finally would take the Coplin jars out and let
them stand for an additional 30 mins at RT. Worked fine for tumors. Any
ideas for joints???
All suggestions would be greatly appreciated.

Igor Deyneko
Infinity Pharmaceuticals
Cambridge, MA 02139
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