Not available outside of the UK & Ireland.

Analysis Note

Protein determined by biuret.

Application

Phosphatase, Alkaline from Escherichia coli has been used:in fluorometric assay to determine its enzyme activityin the dephosphorylating enzyme cocktail for dephosphorylation of peptidesin 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, 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.

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

Caution

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

Physical form

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

Unit Definition

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