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Merck

P6774

Sigma-Aldrich

Phosphatase, Alkaline from bovine intestinal mucosa

buffered aqueous solution, ≥2,000 DEA units/mg protein

Sinónimos:

Alkaline phosphatase, Orthophosphoric-monoester phosphohydrolase (alkaline optimum)

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About This Item

Número de CAS:
Comisión internacional de enzimas:
Número MDL:
Código UNSPSC:
12352204
eCl@ss:
42010105
NACRES:
NA.54

origen biológico

bovine intestinal mucosa

Nivel de calidad

formulario

buffered aqueous solution

actividad específica

≥2,000 DEA units/mg protein

mol peso

dimer ~160 kDa

concentración

5-20 mg/mL

temp. de almacenamiento

2-8°C

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Aplicación

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.
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. It has been conjugated to sheep anti-rabbit IgG and sheep anti-rabbit IgM in immunological assays. 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.

Acciones bioquímicas o fisiológicas

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. Maximal activity with alkaline phosphatase is achieved in the presence of magnesium. 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. Arsenate, cysteine, iodine, inorganic phosphate, pyrophosphate, diisopropyl phosphate, triphenylphosphate, diisopropyl fluorophosphate, and L-phenylalanine are some of the strong inhibitors of alkaline phosphatase.

Propiedades físicas

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.

Definición de unidad

1 DEA-Einheit hydrolysiert 1 μmol 4-Nitrophenyl-Phosphat pro Minute bei pH 9.8 und 37 °C.

Forma física

Solution in 3.0 M NaCl containing 5 mM MgCl2, 0.2 mM ZnCl2, and 30 mM triethanolamine, pH 7.6

Nota de preparación

Affinity purified

Nota de análisis

Package sizes are based on DEA units
Protein determined by biuret.

Pictogramas

Health hazard

Palabra de señalización

Danger

Frases de peligro

Consejos de prudencia

Clasificaciones de peligro

Resp. Sens. 1

Código de clase de almacenamiento

10 - Combustible liquids

Clase de riesgo para el agua (WGK)

WGK 2

Punto de inflamabilidad (°F)

Not applicable

Punto de inflamabilidad (°C)

Not applicable

Equipo de protección personal

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)


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Guy Miranda et al.
Food chemistry: X, 5, 100080-100080 (2020-03-04)
Here we describe a method based on Liquid Chromatography coupled with Mass Spectrometry (LC-MS) that provides an accurate determination of the six main bovine milk proteins, including allelic and splicing variants, as well as isoforms resulting from post-translational modifications, with
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
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
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
Leilei Zhou et al.
Genome biology, 20(1), 156-156 (2019-08-08)
Methylation of nucleotides, notably in the forms of 5-methylcytosine (5mC) in DNA and N6-methyladenosine (m6A) in mRNA, carries important information for gene regulation. 5mC has been elucidated to participate in the regulation of fruit ripening, whereas the function of m6A

Protocolos

Enzymatic Assay of Alkaline Phosphatase, Diethanolamine Assay (EC 3. 1. 3. 1)

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