I have used with great success the alkaline phos method in Bancroft and Cook
(1994) pg 300. This is suitable for paraffin embedded tissues as well. It is
the BCIP-NBT method. I counterstain with Nuclear Fast red for better
contrast. I also use the BCIP-NBT stock solution available from Boehringer
Mannheim (cat # 1681451) use 1 ml of the purchased stock in 37 ml of the
veronal acetate buffer for equal molar substrate to the described procedure.
Good luck, Donna Montague, UAMS Orthopaedic Research Little ROck, AR

Nora wrote:
We would like to look at alk phos in snap frozen rabbit tissue from a bone
harvest chamber.  Can anyone suggest an antibody or substrate that will
work under these conditions?  I appreciate your help!!!!

For those in the US, enjoy your hour gained this weekend.

Nora Fox
Division of Orthopaedic Surgery
Stanford University
Edwards R144, MC 5341
300 Pasteur Drive
Stanford, CA 94305
phone: (650)725-2746
fax:  (650)723-6396