[Histonet] IHC, decalcification preferences

[Amy Porter]

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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