Product Name

Phosphatase, Alkaline from bovine intestinal mucosa

General deion

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.

Synonym

Alkaline phosphatase, Orthophosphoric-monoester phosphohydrolase (alkaline optimum)

Assay

≥2,000 DEA units/mg protein; ≥1.0 mg/mL

CAS Number

9001-78-9

Molecular Weight

dimer mol wt ~160 kDa

MDL number

MFCD00131849

Suitability

BioReagent

l Physical and Chemical Information

Appearance

ammonium sulfate suspension

Physical form

solution in 3.2 M ammonium sulfate, 1 mM MgCl² and 0.1 mM ZnCl², pH 7.0

l Biological Information

Biochem/Physiol Actions

Alkaline phosphatase, from bovine intestinal mucosa, is most stable in the pH range 7.5-9.5. The enzyme has a broad specificity for phosphate esters of alcohols, amines, pyrophosphate, and phenols and it requires zinc, and magnesium or calcium divalent ions for activity. The enzyme has a broad specificity for phosphate esters of alcohols, amines, pyrophosphate, and phenols. It is routinely used to dephosphorylate proteins and nucleic acids.

Application

Alkaline phosphatase can be used to dephosphorylate casein and other proteins. 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..

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, such as casein, 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 P5521 has been used to study GTP hydrolysis .

High specific activity grade recommended for antibody and protein conjugation.

The enzyme from Sigma has been used in the assay of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate. It has also been used for alkaline phosphatase assays using a Nafion^®-modified electrode.

Preparation Note

Affinity purified; This product is prepared from bovine intestinal mucosa. Dilutions should be made in 10 mM Tris-HCl, pH 8.0, 1-5 mM magnesium chloride, 0.1-0.2 mM zinc chloride, and 50% glycerol and stored at 2-8 °C.

Unit Definition

One DEA unit will hydrolyze 1 μmole of 4-nitrophenyl phosphate per minute at pH 9.8 at 37 °C. (One glycine unit is equivalent to ~3 DEA units)

l Storage

Storage temp.

2-8°C

l Precautions and Disclaimer

This product is for R&D use only, not for drug, household, or other uses.

l References

1. http://www.drugbank.ca

2. https://ncit.nci.nih.gov

3. https://www.ncbi.nlm.nih.gov