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The Alkaline Phosphatase (Thermosensitive) produced by our company is a thermally unstable alkaline phosphatase derived from Antarctic microorganisms (TAB5) and recombinantly expressed and purified by E. coli. It can non-specifically catalyze the removal of DNA, RNA, dNTP, and rNTP. 5'or 3'terminal phosphate group. Alkaline Phosphatase is widely used in molecular biology, such as removing phosphorylated ends of DNA and RNA for subsequent use in probe cloning or end labeling; in molecular cloning experiments, dephosphorylation can prevent self-ligation of linearized plasmid DNA; It can also be used to degrade unbound dNTPs in PCR reactions for subsequent sequencing or SNP analysis.
Features: rapid dephosphorylation; unstable to heat, completely inactivated by incubating at 80°C for 2 min.
Source: Escherichia coli strain carrying TAB5 alkaline phosphatase gene; its monomer molecular weight is about 35 kDa;
Enzyme activity definition: the amount of enzyme required to dephosphorylate 1 µg of linearized pUC19 vector by incubating at 37°C for 30 min is defined as one unit of enzyme activity. Dephosphorylation is defined as the inhibition rate of vector self-ligation reaction greater than 95% (measured by E. coli transformation).
Inactivation or inhibition: Heating at 80°C for 2 min can fully inactivate Alkaline Phosphatase. EDTA and other metal ion chelating agents have inhibitory effect on Alkaline Phosphatase
Purity and concentration: SDS-PAGE detection purity ≥95%, 5 U/μL.
Enzyme storage buffer：10 mM Tris-HCl，1 mM MgCl2，0.01 mM ZnCl2，50% Glycerol，pH 7.4。
Enzyme storage buffer: 10 mM Tris-HCl，1 mM MgCl2，0.01 mM ZnCl2，50% Glycerol，pH 7.4。
10×ALP Reaction Buffer：500 mM Bis-Tris-HCl，10 mM MgCl2，1 mM ZnCl2，pH 6.0。
Storage and transportation
Transport with wet ice; storage at -20°C, valid for 12 months.