[Histonet] IHC, decalcification preferences

[Bernie Taupin]

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 <portera <@t> msu.edu>
To: histonet <@t> lists.utsouthwestern.edu; Nicole Collette <collette2 <@t> 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:  portera <@t> msu.edu
Web:  www.humanpathology.msu.edu
----- Original Message ----- From: "Nicole Collette" <collette2 <@t> mail.llnl.gov>
To: <histonet <@t> 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
> 
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