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Key Documents

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:
Enzyme Commission number:
EC Number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54

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.

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

Signal Word

Danger

Hazard Statements

Precautionary Statements

Hazard Classifications

Resp. Sens. 1

Storage Class Code

10 - Combustible liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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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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)
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)
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
Jong Kil Lee et al.
The Journal of experimental medicine, 211(8), 1551-1570 (2014-07-23)
In Alzheimer's disease (AD), abnormal sphingolipid metabolism has been reported, although the pathogenic consequences of these changes have not been fully characterized. We show that acid sphingomyelinase (ASM) is increased in fibroblasts, brain, and/or plasma from patients with AD and

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