P7923
Phosphatase, Alkaline from bovine intestinal mucosa
buffered aqueous glycerol solution, ≥4,000 DEA units/mg protein
Synonyme(s) :
Alkaline phosphatase, Orthophosphoric-monoester phosphohydrolase (alkaline optimum)
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A propos de cet article
Numéro CAS:
UNSPSC Code:
12352204
eCl@ss:
42010105
EC Number:
232-631-4
NACRES:
NA.54
MDL number:
Specific activity:
≥4,000 DEA units/mg protein
Concentration:
≥10.0 mg/mL
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buffered aqueous glycerol solution
Quality Level
specific activity
≥4,000 DEA units/mg protein
mol wt
dimer ~160 kDa
concentration
≥10.0 mg/mL
storage temp.
2-8°C
General description
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.
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.
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 usually conjugated to antibodies and other proteins in immunological techniques such as ELISA, Western blotting, and immunohistochemistry. 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 P7923 has been used in a novel screening strategy for nitrilase-producing strains[1].
High specific activity grade recommended for antibody and protein conjugation.
The enzyme has been used to develop a highly sensitive immunoassay for determining one thousandth of an attomole (1 zeptomole) of alkaline phosphatase. It has been used in an immunoassay of proinsulin.[2] It has also been used to pretreat ISG (immature secretory granule) membranes to study the phosphorylation of proteinacous components in it.[3]
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. It requires zinc, and magnesium or calcium divalent ions for activity[4].
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.
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.[4] Maximal activity with alkaline phosphatase is achieved in the presence of magnesium.[5] 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.[6][7][8] Arsenate, cysteine, iodine, inorganic phosphate, pyrophosphate, diisopropyl phosphate, triphenylphosphate, diisopropyl fluorophosphate, and L-phenylalanine are some of the strong inhibitors of alkaline phosphatase.[6][9][10]
Physical form
Solution in 40% glycerol containing 6 mM Tris/HCl, 6 mM MgCl2 and 0.12 mM ZnCl2, ~pH 7.6
Analysis Note
Package sizes are based on DEA units
Protein determined by biuret.
Other Notes
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)
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Cet article | |||
|---|---|---|---|
| specific activity ≥4,000 DEA units/mg protein | specific activity ≥2,000 DEA units/mg protein | specific activity ≥5,500 DEA units/mg protein | specific activity ≥10 DEA units/mg solid |
| form buffered aqueous glycerol solution | form buffered aqueous solution | form (Solution in 40% glycerol containing 6 mM Tris, 6 mM MgCl2 and 0.12 mM ZnCl2, pH approximately 7.6) | form lyophilized powder |
| concentration ≥10.0 mg/mL | concentration 5-20 mg/mL | concentration ≥10 mg/mL | concentration - |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C | storage temp. −20°C |
| mol wt dimer ~160 kDa | mol wt dimer ~160 kDa | mol wt dimer ~160 kDa | mol wt dimer ~160 kDa |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
signalword
Danger
hcodes
pcodes
Hazard Classifications
Resp. Sens. 1
Classe de stockage
10 - Combustible liquids
wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, type N95 (US)
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Contenu apparenté
QC Methods
D B Cook et al.
Clinical chemistry, 39(6), 965-971 (1993-06-01)
Enzyme amplification has proved to be a highly sensitive quantification technique for immunoassays. We have shown that by using a fluorescent end-point, even more sensitive enzyme amplification assays can be generated than hitherto reported. We describe some general properties of
Zhijie Chen et al.
Proceedings of the National Academy of Sciences of the United States of America, 117(35), 21328-21335 (2020-08-21)
Theoretical and experimental observations that catalysis enhances the diffusion of enzymes have generated exciting implications about nanoscale energy flow, molecular chemotaxis, and self-powered nanomachines. However, contradictory claims on the origin, magnitude, and consequence of this phenomenon continue to arise. To
Yana Konokhova et al.
Skeletal muscle, 6, 10-10 (2016-02-20)
Low mitochondrial content and oxidative capacity are well-established features of locomotor muscle dysfunction, a prevalent and debilitating systemic occurrence in patients with chronic obstructive pulmonary disease (COPD). Although the exact cause is not firmly established, physical inactivity and oxidative stress
Numéro d'article de commerce international
| Référence | GTIN |
|---|---|
| P7923-10KU | 04061834400585 |
| P7923-2KU | 04061834400592 |