Alkaline Phosphatase, Calf Intestinal
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.
- 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.
- Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
- Seeburg, P.H. et al. (1977) Nature 270, 486–94.
- Ullrich, A. et al. (1977) Science 196, 1313–9.
- Meyerowitz, E.M. et al. (1980) Gene 11, 271–82.
- 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 1umol 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).
Alkaline Phosphatase, Calf Intestinal (CIAP) Protocol
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Certificate of AnalysisLookup Certificate of Analysis
-30C TO -10C
For Research Use Only. Not for Use in Diagnostic Procedures.