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DESCRIPTION
Alkaline Phosphatase
catalyszes the removal of 5'-phosphate groups from DNA, RNA and ribo- and
deoxyribonucleoside triphosphates.
SOURCE
Calf intestinal mucosa.
APPLICATIONS
¡Ü Removing 5' phosphates
from DNA or RNA prior to end labeling.
¡Ü Removing 5' phosphates from
fragments of DNA to prevent self-ligation.
UNIT DEFINITION
One unit is defined as
the amount of enzyme required to catalyze the hydrolysis of 1¥ìmol of
4-nitrophenyl- phosphate to 4-nitrophenol in 1 minute at 37¡É.
STANDARD UNIT ASSAY CONDITIONS
1M
diethanolamine (pH9.8), 0.5mM MgCl2, 10.9mM 4-nitrophenylphosphate.
CONCENTRATION: 1-5 unit/¥ìl.
10X REACTION BUFFER
500mM Tris-HCl
(pH9.3), 10mM MgCl2, 1mM ZnCl2, 10mM spermidine.
STORAGE CONDITIONS
10mM Tris-HCl
(pH8.0), 1mM MgCl2, 0.1mM ZnCl2, 50mM KCl and 50% glycerol. Store at
-20¡É.¡¡
REFERENCE
1. Sambrook, J., Fritsch,
E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual,
second ed., Cold Spring Harbor Laboratory, Cold Spring
Harbor. |