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-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala Sent: Wednesday, February 03, 2010 3:49 PM To: Sherwood, Margaret ; histonet@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-boun...@lists.utsouthwestern.edu on behalf of Sherwood, Margaret Sent: Wed 2/3/2010 3:10 PM To: histonet@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) msherw...@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@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet