|Related Categories||3.1.x.x Acting on esters, 3.x.x.x Hydrolases, Alkaline Phosphatase Enzymes, Analytical and Industrial Enzymes, Biochemicals and Reagents,|
|form||buffered aqueous glycerol solution|
|concentration||≥10,000 units/mL in diethanolamine buffer, pH 9.8 at 37°C|
|foreign activity||DNase, RNase, none detected|
|shipped in||wet ice|
|Gene Information||cow ... ALPI(280993)|
Alkaline phosphatase is a phosphomonoesterase that catalyzes the cleavage of terminal phosphates from nucleic acids. The free 5′-OH can be phosphorylated with polynucleotide kinase and γ32P-ATP to produce γ32P-end-labeled nucleic acids. Linearized cloning vectors can be prevented from recircularizing by dephosphorylation with alkaline phosphatase.
Alkaline phosphatase in 5 mM EDTA, pH 8, will be irreversibly heat inactivated at 75 °C for 10 minutes.
One unit will hydrolyze 1.0 μmole of p-nitrophenyl phosphate per min at pH 9.8 at 37 °C.
Solution in 10 mM Tris-HCl, pH 8.2, 50 mM KCl, 1 mM MgCl2, 0.1 mM ZnCl2, 50% (w/v) glycerol.
Calf intestinal alkaline phosphatase is used in molecular biology applications to remove 5′-phosphate groups from nucleic acid molecules to prevent self-ligation of DNA fragments and vectors during recombinant DNA manipulations.
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lyophilized powder, ≥200 units/mg protein (E
BioUltra, buffered aqueous glycerol solution, ≥6,500 DEA units/mg protein
ammonium sulfate suspension, 30-90 units/mg protein (modified Warburg-Christian, in glycine buffer)
1. OBJECTIVE To standardize a procedure for the enzymatic assay of alkaline phosphatase, diethanolamine assay.
Keywords: Enzyme activity, Extinction coefficient, PAGE
Reid, T.W., et al. Enzymes 3, 373-416, (1971)
Sambrook, J., et al. Molecular Cloning: A Laboratory Manual Cold Spring Harbor, NY , (1989), 5.72
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|D6518||Diethylenetriaminepentaacetic acid, ≥99% (titration)|
|P9375||1,10-Phenanthroline monohydrate, reagent grade|
|ED2SS||Ethylenediaminetetraacetic acid disodium salt dihydrate, reagent grade, 98.5-101.5% (titration)|
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|N2770||SIGMAFAST™ p-Nitrophenyl phosphate Tablets, tablet, To prepare 20 mL|
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