[Histonet] IHC, decalcification preferences
Nicole Collette
collette2 <@t> mail.llnl.gov
Wed Apr 1 11:44:03 CDT 2009
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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