alk phos enzyme a la Bancroft modified
1. Prepare 0.2 M tris buffer pH 9.5 (original calls for veronal acetate
2. Prepare 1 M magnesium chloride
3. Obtain NBT/BCIP stock solution (Roche Molecular Biochemicals Cat #
4. Deparaffinize and rehydrate to water.
5. Prepare the staining solution as follows:
37 mL 0.2 M tris buffer
294 uL 1 M MgCl2
1 mL NBT/BCIP stock
filter prior to use into a coplin jar (may use 0.45 um syringe
filter or whatman # 1 filter paper)
6. Place slides into prepared stain and incubate at 37 oC 10 - 30 minutes.
8. counterstain as desired (Nuclear fast red or 0.1 % safranin o works
9. wash, dehydrate, clear and mount.
alkaline phosphatase = dark blue-purple
cytoplasm/nuclei = pink - red
Bancroft and Cook pg 300, 1994
If you have questions please call (501) 603-1239 or email.
Good luck, Donna Montague, M.S.
Physiology/Biophysics and Orthopaedic Surgery
University of Arkansas for Medical Sciences
From: Gerardo Nava [mailto:firstname.lastname@example.org]
Sent: Monday, March 18, 2002 10:39 PM
Subject: Help, please!!!
Dear Dr Donna,
I am looking for the directions for alkaline phosphatase stain, I have found
information that you use the Bancroft & Cook Method found in the Manual of
Histological Techniques and their Diagnostic Application. I was hoping you
could fax me a copy of this protocol.
It would be greatly appreciated. My fax number is 501-5758490.
Thank you so much!!!
University of Arkansas
Room O114, Poultry Science Center
Fayetteville, Arkansas 72701, USA.
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