Description
Alkaline phosphatase (ALP, ALKP, ALPase, Alk Phos) (EC 3.1.3.1) is a hydrolase enzyme responsible for removing phosphate groups from many types of molecules, including nucleotides, proteins, and alkaloids. The process of removing the phosphate group is called dephosphorylation. As the name suggests, alkaline phosphatases are most effective in an alkaline environment. It is sometimes used synonymously as basic phosphatase.
Abbr
ALP, Native (Bovine)
Source
Bovine intestinal mucosa
Applications
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 hist ochemical 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. This product has been used to study the mon oclonal alkaline phosphatase-anti-alkaline phosphatase (APAAP) complex. High specific activity grade recommended for antibody and protein conjugation.
Product Overview
Bovine intestinal alkaline phosphatase is a dimeric, membrane-derived glycoprotein. At least three isoforms exist, which typically possess two N-linked and one or more O-linked glycans per monomer.2 The enzyme requires zinc, and magnesium or calcium divalent ions for activity. Bovine intestinal alkaline phosphatase is a dimeric, membrane-derived glycoprotein. At least three isoforms exist, which typically possess two N-linked and one or more O-linked glycans per monomer.
Form
Type I, lyophilized powder; Type II, aqueous solution, solution in 3.2 M ammonium sulfate, 1 mM MgCl2 and 0.1 mM ZnCl2, pH 7.0; Type III, buffered aqueous solution, Solution in 3.0 M NaCl containing 5 mM MgCl2, 0.2 mM ZnCl2, and 30 mM triethanolamine, pH 7.6; Type IV, Type V, Type VI, buffered aqueous glycerol solution, Solution in 50% glycerol containing 5 mM Tris, 5 mM MgCl2 and 0.1 mM ZnCl2, pH 7.0.
Enzyme Commission Number
EC 3.1.3.1
Activity
Type I, > 10 DEA units/mg solid; Type II, > 2 ,000 DEA units/mg protein; Type III, 2 ,000-4 ,000 DEA units/mg protein; Type IV, > 5,500 DEA units/mg protein; Type V, > 6,500 DEA units/mg protein; Type VI, > 4 ,000 DEA units/mg protein.
Molecular Weight
dimer mol wt ~160 kDa
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)
Warnings
Package sizes are based on DEA units
Pathway
Endochondral Ossification, organism-specific biosystem (from WikiPathways) Folate biosynthesis, organism-specific biosystem (from KEGG) Folate biosynthesis, conserved biosystem (from KEGG) Metabolic pathways, organism-specific biosystem (from KEGG) TNF-alpha NF-kB Signaling Pathway, organism-specific biosystem (from WikiPathways)
Function
The peri-partum characteristics of serum bone-specific alkaline phosphatase (BAP) and urinary deoxypyridinoline (DPD) in dairy cattle are reported. Results indicate that the presence of glycosylphosphatidylinositol increases the stability of alkaline phosphatase against chemical denaturation while it decreases its refolding yield by the artificial chaperone refolding technique. Reliable and reproducible estimates of k (cat) and K (m) from only two or three progress curves were obtained using alkaline phosphatase. GPI-anchored proteins are localized on the outer layer of plasma membranes and clustered in microdomains generally called lipid rafts.
Synonyms
Alkaline phosphatase; ALP; ALKP; ALPase; Alk Phos; EC 3.1.3.1; Alkaline phosphomonoesterase; Glycerophosphatase; Phosphomonoesterase
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