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Merck

P8361

Sigma-Aldrich

Phosphatase, Alkaline bovine

recombinant, expressed in Pichia pastoris, ≥4000 units/mg protein

Sinónimos:

Orthophosphoric-monoester phosphohydrolase (alkaline optimum)

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About This Item

Número de CAS:
Comisión internacional de enzimas:
EC Number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54

recombinant

expressed in Pichia pastoris

form

solid

specific activity

≥4000 units/mg protein

mol wt

apparent mol wt 160 kDa

storage temp.

2-8°C

Application

Alkaline phosphatase is used for conjugation to antibodies and other proteins for ELISA, Western blotting, and histochemical detection. It is routinely used to dephosphorylate proteins and nucleic acids. It may be used for protein labeling when high sensitivity is required. Alkaline phosphatase may be also be used to dephosphorylate the 5′-termini of DNA or RNA to prevent self-ligation. DNA or RNA can also be tagged with radiolabeled phosphate (via T4 polynucleotide kinase) after dephosphorylation with alkaline phosphatase. Product P8361 has been used to dephosphorylate Dishevelled (Dvl) , which is a key component in the Wnt/β-catenin signaling pathway.
An assay, using this ALP enzyme, was developed to assess the cryoprotective activity of two proteins CRP-1 and CRP-2. The enzyme has been used for inhibitory studies of cyclic nucleotide analogs that inhibit prostatic acid phosphatase using a fluorogenic assay. It is commonly used as a "reporter" in detection systems, in which it is conjugated to a protein (antibody, streptavidin, etc.) that specifically recognizes a target molecule. Alkaline phosphatase has also been used to dephosphorylate casein.
The product is manufactured without the use of any animal source materials during fermentation or purification.

Biochem/physiol Actions

Alkaline phosphatase dephosphorylates proteins and nucleic acids.
Alkaline phosphatase is a dimer consisting of two equal subunits. The enzyme is a glycoprotein containing approximately 12% carbohydrate (6% hexoses and 6% other neutral sugars). Each molecule of alkaline phosphatase contains four zinc atoms and four disulfide bridges. Maximal activity with alkaline phosphatase is achieved in the presence of magnesium. It catalyzes the hydrolysis of phosphate monoesters such as p-nitrophenyl phosphate, phenyl phosphate, phenolphthalein phosphate, α-glycerol phosphate, β-glycerol phosphate, 2-phosphorylglycerate, triosephosphate, glucose-6-phosphate, glucose 1-phosphate, fructose 1-phosphate, fructose 6-phosphate, adenosine 5-phosphate adenosine 3-phosphate, phosphoenolpyruvate, and β-nicotinamide adenine dinucleotide phosphate. Arsenate, cysteine, iodine, inorganic phosphate, pyrophosphate, diisopropyl phosphate, triphenylphosphate, diisopropyl fluorophosphate, and L-phenylalanine are some of the strong inhibitors of alkaline phosphatase. The enzyme is most stable in the pH range 7.5-9.5.2. The pH optimum for enzymatic activity is pH 8-10.

Unit Definition

One DEA unit will hydrolyze 1 μmole of 4-nitrophenyl phosphate per minute at pH 9.8 at 37 °C.

pictograms

Health hazard

signalword

Danger

hcodes

Hazard Classifications

Resp. Sens. 1

Storage Class

11 - Combustible Solids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, type N95 (US)


Certificados de análisis (COA)

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The enzymic dephosphorylation of casein fractions in the presence of 0-18 labelled water.
E B KALAN et al.
Archives of biochemistry and biophysics, 85, 273-275 (1959-11-01)
Ondrej Bernatik et al.
The Journal of biological chemistry, 286(12), 10396-10410 (2011-02-03)
Dishevelled (Dvl) is a key component in the Wnt/β-catenin signaling pathway. Dvl can multimerize to form dynamic protein aggregates, which are required for the activation of downstream signaling. Upon pathway activation by Wnts, Dvl becomes phosphorylated to yield phosphorylated and
The activation of alkaline phosphatase by effector molecules. A combined kinetic and hydrodynamic study.
H Neumann et al.
European journal of biochemistry, 109(2), 475-480 (1980-08-01)
Differential staining of phosphoproteins on polyacrylamide gels with a cationic carbocyanine dye.
M R Green et al.
Analytical biochemistry, 56(1), 43-51 (1973-11-01)
Eric S McCoy et al.
Journal of biomolecular screening, 18(4), 481-489 (2012-11-30)
The secretory and transmembrane isoforms of prostatic acid phosphatase (PAP) can dephosphorylate extracellular adenosine 5'-monophosphate (AMP) to adenosine, classifying PAP as an ectonucleotidase. Currently, there are no compounds that inhibit PAP in living cells. To identify small-molecule modulators of PAP

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