P4069

Phosphatase, Alkaline from Escherichia coli

buffered aqueous glycerol solution, 20-50 units/mg protein (in glycine buffer)

• Synonym(s): Orthophosphoric-monoester phosphohydrolase (alkaline optimum)
• CAS Number: 9001-78-9
• UNSPSC Code: 12352204
• NACRES: NA.54
• EC Number: 232-631-4
• MDL number: MFCD00131849
• EC Number: 3.1.3.1 (BRENDA, IUBMB)
• eCl@ss: 42010105
• Specific activity: 20-50 units/mg protein (in glycine buffer)
• Biological source: Escherichia coli

Properties

• biological source: Escherichia coli
• Quality Segment: 200
• form: buffered aqueous glycerol solution
• specific activity: 20-50 units/mg protein (in glycine buffer)
• foreign activity: DNase, RNase, none detected
• storage temp.: −20°C

Description

Application

Phosphatase, Alkaline from Escherichia coli has been used:

• in fluorometric assay to determine its enzyme activity
• in the dephosphorylating enzyme cocktail for dephosphorylation of peptides
• in treating the nerve and HL60 cell lysate for β-hydroxy β-methylglutaryl-Coenzyme A (HMG-CoA) reductase activity

Alkaline phosphatase is used for conjugation to antibodies and other proteins for ELISA, Western blotting, and histochemical detection. It may be used for protein labeling when high sensitivity is required. Product P4069 is provided as a buffered aqueous glycerol solution.

Biochem/physiol Actions

Alkaline phosphatase is a model enzyme for understanding phosphomonoesterase. It is used in various biochemical methods and enzyme linked immunosorbent assay (ELISA).

Alkaline phosphatase, from Escherichia coli, is a dimeric, non-glycosylated protein which mainly reside in the periplasmic space. Three known isoforms exist. The enzyme requires zinc, and is activated by magnesium. E. coli akaline phosphatase has a broad specificity for phosphate esters.

Physical form

Solution in 5 mM Tris-HCl, pH 7.4, 0.5 mM MgCl₂, 0.5 mM ZnCl₂, 50% (w/v) glycerol

Analysis Note

Protein determined by biuret.

Other Notes

One unit will hydrolyze 1.0 μmole of p-nitrophenyl phosphate per min at pH 10.4 at 37 °C.

Disclaimer

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

Safety Information

• pictograms: GHS08
• signalword: Danger
• hcodes: H334
• pcodes: P261 - P284 - P501
• Hazard Classifications: Resp. Sens. 1
• Storage Class: 10 - Combustible liquids
• wgk: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable
• ppe: Eyeshields, Gloves, type N95 (US)