Alkaline Phosphatase (Calf Intestine)

Calf intestine derived phosphatase which catalyzes the hydrolysis of phosphate monoesters.

Description:

• Has higher turnover rate and milder conditions of inactivation than bacterial enzyme (1).
• Can be used to remove 5'-phosphates from DNA or RNA prior to 5'-end labeling (1).
• Used to remove 5'-phosphates from linearized vector molecules to prevent self-ligation of the vector during cloning procedures (1).
• Applicable to dephosphorylation of proteins.

Unit Definition: One unit is the amount of enzyme required to hydrolyze 1 µmol of p-nitrophenylphosphate to p-nitrophenol in 1 min at 37°C in a buffer of 1 M diethanolamine, 10 mM p-nitrophenylophosphate, 0.25 mM MgCl2 (pH 9.8) (2).

Storage Conditions: Store at –20°C.

Storage Buffer: 10 mM Tris-HCl (pH 8.0 at 22°C), 0.05 M NaCl and 50% (v/v) glycerol.

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

References:

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

2. Mosser, E., Boll, M. and Pfleiderer, G. (1980) Hoppe-Seyler’s Z. Physiol. Chem. 361, 543-549.