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Alkaline Phosphatase, Calf Intestinal (CIAP)

CIAP catalyzes the Hydrolysis of 5´ Phosphate Groups from DNA, RNA, and r- and dNTPs

  • Available at high concentration; Cat.# M2825 contains 1,000 units of CIAP at 20u/μl
  • Qualified for blue/white cloning; our assay provides a higher level of quality control for enzymes used in cloning applications

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Description

Size

concentration: 1u/µl
Size: 1,000u
Catalog number: M1821

concentration: 20u/µl
Size: 1,000u
Catalog number: M2825

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).

Protocols

Complete Protocol

Alkaline Phosphatase, Calf Intestinal (CIAP) Protocol


[ts-heading h2=”Specifications” txt_align=”center” css_animation=”fadeInUp”][/ts-heading]

What’s in the box?
concentration: 1u/µl
Size: 1,000u

[wpdatatable id=391 table_view=regular]

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

 

What’s in the box?
concentration: 20u/µl
Size: 1,000u

[wpdatatable id=392 table_view=regular]

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB