biological source
Escherichia coli
Quality Level
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
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Application
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.
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
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.
Unit Definition
One unit will hydrolyze 1.0 μmole of p-nitrophenyl phosphate per min at pH 10.4 at 37 °C.
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.
Disclaimer
Alkaline phosphatase in 5 mM EDTA, pH 8, will be irreversibly heat inactivated at 75°C for 10 minutes.
signalword
Danger
hcodes
pcodes
Hazard Classifications
Resp. Sens. 1
Storage Class
10 - Combustible liquids
wgk_germany
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, type N95 (US)
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