I, personally, think EDTA SUCKS. a) it takes too long b) its not safe - http://en.wikipedia.org/wiki/EDTA#Toxicity_and_environmental_considerations c) its to old-fashioned.
Me, I'm a big proponent of Formic Acid decal. Holla off-list if you want me to hit you up with a protocol, yo. ________________________________ From: Amy Porter <port...@msu.edu> To: histonet@lists.utsouthwestern.edu; Nicole Collette <collet...@mail.llnl.gov> Sent: Wednesday, April 1, 2009 12:56:43 PM Subject: Re: [Histonet] IHC, decalcification preferences All the mouse femurs and tibias done in our laboratory are decaled in 14% EDTA and we have a high success rate with our immunohistochemistry and Tunel staining. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 884-5026 Fax: (517) 432-1368 Email: port...@msu.edu Web: www.humanpathology.msu.edu ----- Original Message ----- From: "Nicole Collette" <collet...@mail.llnl.gov> To: <histonet@lists.utsouthwestern.edu> Sent: Wednesday, April 01, 2009 12:44 PM Subject: [Histonet] IHC, decalcification preferences > Hello, all, > > I am going to be doing some IHC on adult mouse bone, and since I'm new to all > this antigen preservation stuff, was wondering the best way to decalcify? > Normally, I use high percentage EDTA (17%) and determine end point by weight > loss/weight gain. (up until now have been using this protocol for LacZ > stains, works like a charm! ) We are comparing several lines of mice with > bone mineral density differences, and endpoint is very phenotype and > age-dependent. It takes a long time, but the results are worth the wait. I > also have Cal-Rite, which is a formaldehyde/formic acid decalcifier, and > obviously works much faster. I am leaning toward the EDTA, since it seems to > work well for the LacZ, which is sensitive to lots of other processes, but if > there's a reason not to, please let me know. Thanks for all your help and > support, I have so far been given great advice from this listserv. > > Sincerely, > Nicole Collette > LLNL/UCB > > _______________________________________________ > 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
