LacZ activity in decalcified bone

From:Ryan Blair Rountree

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

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

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