Alkaline Phosphatase, Calf Intestinal (CIAP)

Alkaline Phosphatase Product Details

Alkaline Phosphatase, Calf Intestinal (CIAP), catalyzes the hydrolysis of 5´-phosphate groups from nucleic acids and nucleotide triphosphates. This enzyme is used to prevent recircularization and religation of linearized cloning vector DNA by removing phosphate groups from both 5´-termini and may also be used for the dephosphorylation of 5´ phosphorylated ends of DNA or RNA for subsequent labeling with [³²P]ATP and T4 Polynucleotide Kinase. CIAP is active on 5´ overhangs, 5´ recessed and blunt ends. CIAP is provided with 10X Reaction Buffer: 0.5M Tris-HCl (pH 9.3 at 25°C), 10mM MgCl₂, 1mM ZnCl₂, 10mM spermidine.

Applications

• Preventing religation of linearized cloning vehicle DNA by removing phosphate groups from both 5´ termini.
• Removing 5´ phosphate groups prior to end-labeling with T4 Polynucleotide Kinase.

References

1. Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
2. Seeburg, P.H. et al. (1977) Nature 270, 486–94.
3. Ullrich, A. et al. (1977) Science 196, 1313–9.
4. Meyerowitz, E.M. et al. (1980) Gene 11, 271–82.
5. Grosveld, F.G. et al. (1981) Gene 13, 227–37.

Storage Buffer: 10mM Tris-HCl (pH 8.0), 1mM MgCl₂, 0.1mM ZnCl₂, 50mM KCl, 50% (v/v) glycerol.

Source: Calf intestinal mucosa.

QC Tests: Activity, endonuclease, DNase, RNase, blue/white cloning assay.

Unit Definition: One unit is defined as the amount of enzyme required to catalyze the hydrolysis of 1μmol of 4-nitrophenyl phosphate per minute at 37°C in 1M diethanolamine, 10.9mM 4-nitrophenyl phosphate, 0.5mM MgCl₂ (pH 9.8 at 25°C).