Alkaline Phosphatase (E. coli)

Bacterial phosphatase catalyzes the hydrolysis of phosphate esters, including those present in nucleic acids and nucleotides.

Description:

• More thermal stable than Calf Intestine Alkaline Phosphatase (CIAP).
• Optimal incubation temperature is approximately 60°C, however the enzyme remains active from 20°C to 80°C.
• Resistant to chemical changes and active over a broad range of buffer conditions.
• Can be used to remove 5’-phosphates from DNA or RNA prior to 5’-end labeling (1).
• Works to remove 5’-phosphates from linearized vector molecules to prevent self-ligation of the vector during cloning procedures (1).
• Ideal for diagnostic immunoassays and immunodetection of proteins and nucleic acids following blotting experiments (1).

Unit Definition: One unit is the amount of enzyme required to hydrolyze 1 mol of p-nitrophenylphosphate in 1 min at 37°C in a buffer of 1 M diethanolamine, 10 mM p-nitrophenylphosphate, 0.25 mM MgCl₂ (pH 9.8).

Storage Buffer: 20 mM Tris-HCl (pH 7.0 at 22°C), 5 mM potassium phosphate, 100 mM KCl, 0.1 mM MgCl₂, 0.1 mM ZnCl₂ and stabilizers.

Storage Conditions: Store at -20°C.

Quality Control: All preparations are assayed for contaminating endonuclease and nonspecific RNase and single- and double-stranded DNase activities.

References:

1. Sambrook, J. et al. (1989) Molecular cloning: A laboratory Manual, second edition, pp. 1.56, 5.72 Cold Spring Harbor, New York.