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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CN¥483.36
CN¥483.36
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1 ML
CN¥483.36
About This Item
CAS Number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54
CN¥483.36
Available to ship onApril 27, 2025Details
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form
aqueous solution
Quality Level
specific activity
≥2,000 DEA units/mg protein
mol wt
dimer ~160 kDa
concentration
≥1.0 mg/mL
storage temp.
2-8°C
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Related Categories
1 of 4
This Item | F8636 | F9378 | F2637 |
|---|---|---|---|
| technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable | technique(s) - | technique(s) - |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
| product line BioReagent | product line BioXtra | product line - | product line BioXtra |
| type Type 400-DL | type - | type Type 400-DL | type - |
organelle preparation, centrifugation gradients | organelle preparation, centrifugation gradients | - | - |
General description
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..
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 [8].
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 properties
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.
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)
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.[11]
Analysis Note
Package sizes are based on DEA units
Protein determined by biuret.
Legal Information
Nafion is a trademark of The Chemours Company FC, LLC
inhibitor
Product No.
Description
Pricing
Storage Class Code
10 - Combustible liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
Regulatory Information
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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
Alkaline phosphatase (EC 3.1.3.1).
M J Maunders
Methods in molecular biology (Clifton, N.J.), 16, 331-341 (1993-01-01)
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)
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
Articles
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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