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P4069

Sigma-Aldrich

Phosphatase, Alkaline from Escherichia coli

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

Synonym(s):

Orthophosphoric-monoester phosphohydrolase (alkaline optimum)

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About This Item

CAS Number:
9001-78-9
Enzyme Commission number:
3.1.3.1 (BRENDA, IUBMB)
EC Number:
232-631-4
MDL number:
MFCD00131849
UNSPSC Code:
12352204
NACRES:
NA.54

biological source

Escherichia coli

Quality Level

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

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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.

Caution

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

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.

pictograms

Health hazard
GHS08

signalword

Danger

hcodes

H334

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)

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Sensitive fluorogenic substrate for alkaline phosphatase
Levine MN and Raines RT
Analytical Biochemistry, 418(2), 247-252 (2011)
The prion protein and cellular cholesterol homeostasis
Diomede L, et al.
Neurobiology of Lipids, 1(2), 8-14 (2002)
Phospho-iTRAQ data article: Assessing isobaric labels for the large-scale study of phosphopeptide stoichiometry
Glibert P, et al.
Data in Brief, 4(2), 60-65 (2015)
Lionel M Igaz et al.
The American journal of pathology, 173(1), 182-194 (2008-06-07)
TAR DNA-binding protein (TDP-43) has been recently described as a major pathological protein in both frontotemporal dementia with ubiquitin-positive inclusions (FTLD-U) and amyotrophic lateral sclerosis. However, little is known about the relative abundance and distribution of different pathological TDP-43 species
Yixing Cheng et al.
Journal of biomedical nanotechnology, 10(2), 287-298 (2014-04-18)
Scaffold architecture, surface topography, biochemical and mechanical cues have been shown to significantly improve cellular events and in vivo tissue regeneration. Specifically electrospun nanofiber matrices have gained tremendous interest due to their intrinsic structural resemblance to native tissue extracellular matrix
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