Hello again, and thanks to everyone that has answered other questions from
me!

Does anyone have a protocol for decalcifying tissue that maintains LacZ
activity?  Especially in mouse limbs?  I get really strong lacZ activity
cryosectioning calcified young mouse limbs (up to 2 weeks old) by fixing
in 4% paraformaldehyde for 4 hours, followed with 15% sucrose for an hour,
then 30% sucrose overnight and OCT freezing.  This time I simply added a
step after fixation of 10% EDTA pH7.4 for 3 days, followed by the sucrose
and OCT freezing (ALL steps done at 4 degrees C).  But the lacZ activity
was totally dead.  Any suggestions?  THANK YOU!!

Ryan

Ryan Rountree
Department of Developmental Biology
B301, Beckman center
279 Campus Drive
Stanford, CA 94305
(650) 725-7599
Fax: (650) 725-7739