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Bacterial Alkaline Phosphatase

Description
E. coli alkaline phosphatase, which is a dimeric, zinc and magnesium containing
protein with a molecular weight of 80 kDa, is a valuable reagent for removal of
terminal monoesterified phosphate from both ribo- and deoxyribo-oligonucleotide.

Properties
Clinical for in vitro diagnostic use
IVD for in vitro diagnostic use
Storage Temp. Stable at 2-8 ℃ for at least 6-12 months (do not freeze)

Speificity
Alkaline Phosphatase catalyzed the hydrolysis of numerous phosphate esters
including esters of primary and secondary alcohols, amines, phenols, cyclic
alcohols, and saccarides. Phosphodiesters can not be hydrolyzed. The enzyme
hydrolyzes inorganic pyrophosphate. The enzymatic kinetics depends on factors
including buffer, pH, concentration and purity of enzyme.

Applications
Alkaline Phosphatase is coupled to other proteins via its carbohydrate or amino
groups. For conjugation of AP with IgG a ration of 2.5:1 is recommended. The
application of the conguated enzyme varies from ELISAs to Western blot, to
Microarrays. The enzyme solution in NaCl can be used directly for coupling
without further dialysis.

Specific Activity ≧ 6000 U/mg
The enzyme activity is determined at 25 ℃ in 1.5 M Tris-HCl buffer, pH8.0,
with p-nitrophenylphosphate (PNP) as substrate. The E. coli enzyme binds
phosphate tightly over a wide range of pH forming complexes that may be
intermediate in the hydrolytic action.
Substrates for Alkaline Phosphatase

ELISA:
4-Nitrophenyl-phosphate [pNPP]

Blotting and Immunohisto/cytochemistry:
5-Bromo-4-Chloro-3-indolyl phosphate [BCIP]

Labeling and Detection of Nucleic Acids:
CDP-Star

Purity (HPLC)
≧ 95%

Optimum pH
8.0

Protein Concentration
20 mg/ml

General Instructions for Labeling Immunoglobulines with
Enzymes Are in the Flollowing Literatures:
Immunochemistry (1971) 8, 871.
Ann. Clin. Biochem. (1979) 16, 221.
J. Immunol. (1979) 123, 1548.