[Histonet] TRAP on bones

[Patsy Ruegg]

Decalcification with formic acid and formalin fixation will destroy the TRAP
enzyme.  I always use undecalcified bone fixed in cold methanol and embedded
in GMA to demostrate osteoclasts.  Cold Methanol will also best preserve
fluorochrome labels which may be soluable in aqueous fixatives.  Leslie, I
do contract GMA work.  You can visit my website at www.ihctech.net 
Patsy Ruegg 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gayle Callis
Sent: Friday, May 06, 2005 10:08 AM
To: Dixon, Leslie E.; Histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] TRAP on bones

EDTA decalcification is preferred decalcifier.  Formic acid is more than
likely killing the enzyme, rendering it unstainable.  I have a method from
Hermina Borgerink, if you want it.


At 10:05 AM 5/6/2005, you wrote:
>Good morning all,
>
>I have been trying to stain formic acid/EDTA decalcified formalin fixed 
>mouse femurs for osteoclasts with the TRAP stain.  I have had little 
>success.  Does anyone have a protocol that I might be able to try?  Any 
>suggestions would be greatly appreciated.  I have been using the Sigma 
>kit #387A thus far.
>
>Thanks,
>Leslie Dixon
>Mayo Clinic Scottsdale Research Histology
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)



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