NEB, M0289L, Antarctic Phosphatase
Antarctic Phosphatase (AnP) is a heat labile alkaline phosphatase purified from a recombinant source.
Catalog Number: M0289L
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Pack Size0289 L
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| <b>Unit Definition</b> One unit is defined as the amount of enzyme that will dephosphorylate 1 µg of pUC19 vector DNA cut with a restriction enzyme generating 5´ recessed ends in 30 minutes at 37°C. Dephosphorylation is defined as > 95% inhibition of recircularization in a self-ligation reaction and is measured by transformation into E. coli . <b>Reaction Conditions</b> 1X Antarctic Phosphatase Reaction Buffer Incubate at 37°C 1X Antarctic Phosphatase Reaction Buffer 50 mM Bis-Tris-Propane-HCl 1 mM MgCl2 0.1 mM ZnCl2 (pH 6 @ 25°C) <b>Storage Buffer</b> 10 mM Tris-HCl 1 mM MgCl2 50% Glycerol 0.01 mM ZnCl2 pH 7.4 @ 25°C <b>Heat Inactivation</b> 80°C for 2 minutes <b>Molecular Weights</b> Apparent | 35 kDa Theoretical: 69 kDa <b>Unit Assay Conditions</b> Vector DNA is dephosphorylated in restriction endonuclease buffer supplemented with Antarctic Phosphatase Reaction Buffer. Ligation is performed with 50 ng of vector using the NEB Quick Ligation Kit (NEB #M2200). |
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