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Cat. No.: APT-1k (for 1KU)

Cat. No.: APT-5k (for 5KU)

Description

Alkaline Phosphatase (Thermosensitive) is a heat-labile alkaline phosphatase derived from Antarctic microorganisms (TAB5), recombinantly expressed and purified from Escherichia coli. This enzyme non-specifically catalyzes the removal of 5' or 3' phosphate groups from DNA, RNA, dNTPs, or rNTPs. Alkaline Phosphatase is widely used in molecular biology, for instance, to remove phosphorylated ends from DNA and RNA, facilitating subsequent cloning or end-labeling for probes. In molecular cloning experiments, dephosphorylation prevents the self-ligation of linearized plasmid DNA. It is also employed to degrade unincorporated dNTPs in PCR reactions for subsequent sequencing or SNP analysis.

Features

• Rapid dephosphorylation
• Heat-labile, fully inactivated by incubation at 80°C for 2 minutes

Source

E. coli strain carrying the TAB5 alkaline phosphatase gene; the monomer has a molecular weight of approximately 35 kDa

Activity Definition

One unit of enzyme activity is defined as the amount of enzyme required to dephosphorylate 1 µg of linearized pUC19 vector in 30 minutes at 37°C. Dephosphorylation is defined as achieving a self-ligation inhibition rate of over 95% (as measured by E. coli transformation).

Inactivation or Inhibition

Alkaline Phosphatase is fully inactivated by heating at 80°C for 2 minutes. EDTA and other metal ion chelators inhibit Alkaline Phosphatase activity.

Purity and Concentration

Purity ≥95% as determined by SDS-PAGE, 5 U/μL.

Phosphatases:

• Alkaline Phosphatase (Thermosensitive)
• Antarctic Phosphatase (Thermosensitive)
• Shrimp Alkaline Phosphatase
• Inorganic Yeast Pyrophosphatase
• Inorganic E.coli Pyrophosphatase
• Thermostable Inorganic Pyrophosphatase

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