P6774
Phosphatase, Alkaline from bovine intestinal mucosa
buffered aqueous solution, ≥2,000 DEA units/mg protein
Synonym(s):
Alkaline phosphatase, Orthophosphoric-monoester phosphohydrolase (alkaline optimum)
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About This Item
CAS Number:
UNSPSC Code:
12352204
eCl@ss:
42010105
NACRES:
NA.54
MDL number:
Specific activity:
≥2,000 DEA units/mg protein
Biological source:
bovine intestinal mucosa
Concentration:
5-20 mg/mL
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biological source
bovine intestinal mucosa
form
buffered aqueous solution
specific activity
≥2,000 DEA units/mg protein
mol wt
dimer ~160 kDa
concentration
5-20 mg/mL
storage temp.
2-8°C
Quality Level
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Related Categories
1 of 4
This Item | |||
|---|---|---|---|
| specific activity ≥2,000 DEA units/mg protein | specific activity ≥4,000 DEA units/mg protein | specific activity ≥2,000 DEA units/mg protein | specific activity ≥5,500 DEA units/mg protein |
| biological source bovine intestinal mucosa | biological source - | biological source - | biological source - |
| concentration 5-20 mg/mL | concentration ≥10.0 mg/mL | concentration ≥1.0 mg/mL | concentration ≥10 mg/mL |
| form buffered aqueous solution | form buffered aqueous glycerol solution | form aqueous solution | form (Solution in 40% glycerol containing 6 mM Tris, 6 mM MgCl2 and 0.12 mM ZnCl2, pH approximately 7.6) |
| mol wt dimer ~160 kDa | mol wt dimer ~160 kDa | mol wt dimer ~160 kDa | mol wt dimer ~160 kDa |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C | 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 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 P6774 has been used during preparation of brain lysate[1].
High specific activity grade recommended for antibody and protein conjugation.
The enzyme from Sigma has been used in the preparation of AP-BGG (alkaline phosphatase-bovine gamma globulin) conjugates.[2] It has been conjugated to sheep anti-rabbit IgG and sheep anti-rabbit IgM in immunological assays.[3] It has also been used to prepare double-enzyme conjugates with HRP for the simultaneous detection of three different intracellular Ig determinants in a single tissue section. [4]
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.
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.[5] Maximal activity with alkaline phosphatase is achieved in the presence of magnesium.[6] 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.[7][8][9] Arsenate, cysteine, iodine, inorganic phosphate, pyrophosphate, diisopropyl phosphate, triphenylphosphate, diisopropyl fluorophosphate, and L-phenylalanine are some of the strong inhibitors of alkaline phosphatase.[7][10][11]
Physical form
Solution in 3.0 M NaCl containing 5 mM MgCl2, 0.2 mM ZnCl2, and 30 mM triethanolamine, pH 7.6
Preparation Note
Affinity purified
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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flash_point_f
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N Van Rooijen et al.
Immunology, 51(3), 417-421 (1984-03-01)
Mice were injected intravenously with 2 mg of a bovine gamma globulin-penicilloyl (BGG-Pen) conjugate. Cells producing specific antibodies against the protein carrier bovine gamma globulin (BGG) and cells producing specific antibodies against the hapten penicilloyl (Pen) could be distinguished simultaneously
E Claassen et al.
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 34(4), 423-428 (1986-04-01)
A new double-enzyme conjugate was synthesized by coupling alkaline phosphatase (AP) to horseradish peroxidase (HRP). After AP (blue) and subsequent HRP (red) cytochemistry, this new conjugate produced a stable intermediate-colored (violet) product. By coupling this double-enzyme conjugate to an antigen
N Van Rooijen et al.
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 32(7), 677-680 (1984-07-01)
Rabbits were primed intravenously with human serum albumin (HSA) and boosted with the same antigen 2 months later. Cells producing specific antibodies against HSA could be detected in vivo and it could be determined whether or not they belonged to
Rob U Onyenwoke et al.
Molecular biology of the cell, 23(2), 381-389 (2011-11-25)
AMP-activated protein kinase (AMPK) is a key energy sensor that regulates metabolism to maintain cellular energy balance. AMPK activation has also been proposed to mimic benefits of caloric restriction and exercise. Therefore, identifying downstream AMPK targets could elucidate new mechanisms
Daniel Diaz-de-Quijano et al.
Heliyon, 6(11), e05582-e05582 (2020-12-10)
Extracellular enzyme activities (EEA) are crucial components of microbial food web interactions and biogeochemical cycles in aquatic ecosystems. They also represent relevant biological traits in the ecophysiology of phytoplankton and other components of microbial plankton. To assess species-specific and (sub-)population-level
Protocols
Enzymatic Assay of Alkaline Phosphatase, Diethanolamine Assay (EC 3. 1. 3. 1)
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