P5521
Phosphatase, Alkaline from bovine intestinal mucosa
≥2,000 DEA units/mg protein
Synonym(s):
Alkaline phosphatase, Orthophosphoric-monoester phosphohydrolase (alkaline optimum)
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1000 UNITS
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About This Item
CAS Number:
UNSPSC Code:
12352204
NACRES:
NA.54
MDL number:
eCl@ss:
42010105
Specific activity:
≥2,000 DEA units/mg protein
Concentration:
≥1.0 mg/mL
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form
aqueous solution
specific activity
≥2,000 DEA units/mg protein
mol wt
dimer ~160 kDa
concentration
≥1.0 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 ≥5,500 DEA units/mg protein | specific activity ≥4,000 DEA units/mg protein | specific activity ≥10 DEA units/mg solid |
| form aqueous solution | form (Solution in 40% glycerol containing 6 mM Tris, 6 mM MgCl2 and 0.12 mM ZnCl2, pH approximately 7.6) | form buffered aqueous glycerol solution | form lyophilized powder |
| concentration ≥1.0 mg/mL | concentration ≥10 mg/mL | concentration ≥10.0 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 |
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.
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.[1] Maximal activity with alkaline phosphatase is achieved in the presence of magnesium.[2] 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.[3][4][5] Arsenate, cysteine, iodine, inorganic phosphate, pyrophosphate, diisopropyl phosphate, triphenylphosphate, diisopropyl fluorophosphate, and L-phenylalanine are some of the strong inhibitors of alkaline phosphatase.[3][6][7]
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..
The enzyme from Sigma has been used in the assay of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate.[8] It has also been used for alkaline phosphatase assays using a Nafion™-modified electrode.
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 [9].
High specific activity grade recommended for antibody and protein conjugation.
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.
Physical form
solution in 3.2 M ammonium sulfate, 1 mM MgCl2 and 0.1 mM ZnCl2, pH 7.0
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.[10]
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)
Legal Information
Nafion is a trademark of The Chemours Company FC, LLC
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Storage Class
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wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, type N95 (US)
Regulatory Listings
Regulatory Listings are mainly provided for chemical products. Only limited information can be provided here for non-chemical products. No entry means none of the components are listed. It is the user’s obligation to ensure the safe and legal use of the product.
P5521-VAR: + P5521-100KU: + P5521-BULK: + P5521-50KU: + P5521-1KU: + P5521-10KU: + P5521-2KU:
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Alkaline phosphatase (EC 3.1.3.1).
M J Maunders
Methods in molecular biology (Clifton, N.J.), 16, 331-341 (1993-01-01)
P T Hawkins et al.
The Biochemical journal, 218(3), 785-793 (1984-03-15)
We have developed methods that yield estimates of the 32P content of each of the individual phosphate groups of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate, thus extending the information available from studies of the labelling of these lipids in intact cells
Determination of alkaline phosphatase using a Nafion?-modified electrode.
Le Gal La Salle A, et al.
Journal of Electroanalytical Chemistry, 379(1), 281-291 (1994)
B A Ponder et al.
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 31(7), 911-919 (1983-07-01)
The immunohistological demonstration of H2 antigens in cryostat sections of a wide variety of mouse tissues is reported. The purpose in developing the method was to use H2 antigens as cellular markers in studies of mouse chimeras. Monoclonal anti-H2 antibodies
Xinlin Du et al.
Proceedings of the National Academy of Sciences of the United States of America, 101(24), 8858-8863 (2004-06-05)
A remote labeling method has been developed to determine (18)O kinetic isotope effects (KIEs) in Ras-catalyzed GTP hydrolysis. Substrate mixtures consist of (13)C-depleted GTP and [(18)O,(13)C]GTP that contains (18)O at phosphoryl positions of mechanistic interest and (13)C at all carbon
Articles
硫酸アンモニウム懸濁液として提供される酵素の使用方法
Instructions for working with enzymes supplied as ammonium sulfate suspensions
Protocols
Enzymatic Assay of Alkaline Phosphatase, Diethanolamine Assay (EC 3. 1. 3. 1)
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