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Key Documents

P8361

Sigma-Aldrich

Phosphatase, Alkaline bovine

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

Sinonimo/i:

Orthophosphoric-monoester phosphohydrolase (alkaline optimum)

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About This Item

Numero CAS:
Classificazione EC (Enzyme Commission):
Numero CE:
Numero MDL:
Codice UNSPSC:
12352204
NACRES:
NA.54

Ricombinante

expressed in Pichia pastoris

Forma fisica

solid

Attività specifica

≥4000 units/mg protein

PM

apparent mol wt 160 kDa

Temperatura di conservazione

2-8°C

Applicazioni

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.

Azioni biochim/fisiol

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.

Definizione di unità

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

Pittogrammi

Health hazard

Avvertenze

Danger

Indicazioni di pericolo

Consigli di prudenza

Classi di pericolo

Resp. Sens. 1

Codice della classe di stoccaggio

11 - Combustible Solids

Classe di pericolosità dell'acqua (WGK)

WGK 1

Punto d’infiammabilità (°F)

Not applicable

Punto d’infiammabilità (°C)

Not applicable

Dispositivi di protezione individuale

Eyeshields, Gloves, type N95 (US)


Certificati d'analisi (COA)

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I clienti hanno visto anche

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)
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
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)

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