[Histonet] TRAP assay - acid phosphatase

[Patsy Ruegg]

I use TRAP enzyme histochemical assay from Sigma for osteoclasts on formalin
fixed EDTA decaled Bone paraffin sections, like Liz mentioned as well as
undecalcified frozen bone sections cut with a permanent tungsten carbide
blade in the cryostat using the Instrumedics tape transfer system.  My
frozen bone sections are also formalin fixed and sucrose infiltrated before
freezing.

I do not fix the frozen sections because they are already fixed in formalin.
One thing I do with that assay that may not be mentioned in the protocol
which is mainly for smears, is that I incubate at 37dc for at least an hour
and sometimes for 4 hours or more on paraffin or frozen tape sections.

Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net 
www.ihcrg.org


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Liz Chlipala
Sent: Wednesday, February 03, 2010 3:49 PM
To: Sherwood, Margaret ; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] TRAP assay - acid phosphatase

We have used it on FFPE EDTA decaled mouse bone, with a little bit of
fiddling with the protocol it works pretty good
 
Liz

________________________________

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Sherwood,
Margaret 
Sent: Wed 2/3/2010 3:10 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] TRAP assay - acid phosphatase 



Has anybody  used the TRAP assay kit from Sigma for bone?  We want to use it
on
mouse tibia for IHC.

If so, 1) can you do it on paraffin and/or frozen sections?
       2) if you do it on frozens, do you decalcify first?
       3) what type of blade do you use for sectioning on the cryostat? 
       3) what procedure do you follow after sectioning:  do you fix slides
(i.e. acetone, etc.)

Thank you.

Peggy

Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDW 214)
Massachusetts General Hospital
55 Fruit Street
Boston, Massachusetts 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood <@t> partners.org



The information in this e-mail is intended only for the person to whom it is
addressed. If you believe this e-mail was sent to you in error and the
e-mail
contains patient information, please contact the Partners Compliance
HelpLine at
http://www.partners.org/complianceline . If the e-mail was sent to you in
error
but does not contain patient information, please contact the sender and
properly
dispose of the e-mail.


_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet