P4978

Phosphatase, Alkaline from calf intestine

buffered aqueous glycerol solution

• Synonym(s): CIAP, CIP, Orthophosphoric-monoester phosphohydrolase (alkaline optimum)
• CAS Number: 9001-78-9
• Enzyme Commission number: 3.1.3.1 (BRENDA, IUBMB)
• MDL number: MFCD00131849
• UNSPSC Code: 12352204
• eCl@ss: 42010105
• NACRES: NA.53

Properties

• grade: for molecular biology
• form: buffered aqueous glycerol solution
• mol wt: ~80 kDa
• concentration: ≥10,000 units/mL
• UniProt accession no.: P19111
• foreign activity: DNase, RNase, none detected
• shipped in: wet ice
• storage temp.: −20°C
• Gene Information: cow ... ALPI(280993)

Description

General description

Alkaline phosphatase is a phosphomonoesterase that catalyzes the cleavage of terminal phosphates from nucleic acids. The free 5′-OH can be phosphorylated with polynucleotide kinase and γ32P-ATP to produce γ32P-end-labeled nucleic acids. Linearized cloning vectors can be prevented from recircularizing by dephosphorylation with alkaline phosphatase.

Application

Commonly used to remove the 5′-terminal phosphate from nucleic acids during molecular cloning reactions.

Components

Alkaline phosphatase is provided in a solution of 10 mM Tris-HCl (pH 8.2), 50 mM KCl, 1 mM MgCl₂, and 0.1 mM ZnCl₂, in 50% (w/v) glycerol. 10X CIP Buffer, Product Number C3225, is included.

Unit Definition

One unit will hydrolyze 1 μmole of p-nitrophenyl phosphate per min at 37 °C.

Other Notes

Alkaline phosphatase in 5 mM EDTA (pH 8) will be irreversibly heat inactivated at 75 °C for 10 minutes.

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable