Alkaline phosphatase catalyzes the removal of 5' phosphate groups from DNA, RNA and ribo- and deoxyribonucleoside triphosphates.
Applications:
- removing 5'and 3'phosphoryl groups from nucleic acids;
- preparing templates for 5'end labeling;
- preventing fragments from self ligating;
- dephosphorylation of proteins
Unit
One unit is the amount of enzyme that hydrolyzes 1 umol of p-nitrophenylphosphate to p- nitrophenol in a total reaction volume of 1 ml in 1 min at 37oC.
Unit Assay Conditions: 1 M diethanolamine-HCl (pH 9.8), 0.5 mM MgCl2 and 10 mM p-nitrophenylphosphate. These conditions are only used for quantitating enzyme activity.
Reaction buffer
SE-buffer O, (50 mM Tris-HCl (pH 7.6 at 25oC); 10 mM MgCl2; 100 mM NaCl; 1 mM DTT.)
Optimal temperature
37oC
Storage conditions
50 mM KCl, 10 mM Tris-HCl (pH 8.2), 1 mM MgCl2, 0.1 mM ZnCl2, 50% glycerol. Store at -20oC
Quality control
Purified free of contaminating exonuclease, endonuclease, ribonuclease activities
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