P6774
Fosfatasi, alcalina
buffered aqueous solution, ≥2,000 DEA units/mg protein
Sinonimo/i:
Fosfatasi alcalina, Fosfoidrolasi di monoesteri ortofosforici (alcalino ottimale)
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About This Item
Numero CAS:
Numero MDL:
Codice UNSPSC:
12352204
eCl@ss:
42010105
NACRES:
NA.54
Prodotti consigliati
Origine biologica
bovine intestinal mucosa
Forma fisica
buffered aqueous solution
Attività specifica
≥2,000 DEA units/mg protein
PM
dimer ~160 kDa
Concentrazione
5-20 mg/mL
Temperatura di conservazione
2-8°C
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Categorie correlate
Applicazioni
Grado di attività altamente specifico consigliato per la coniugazione di anticorpi e proteine.
La fosfatasi alcalina può essere utilizzata per defosforilare la caseina e altre proteine. La fosfatasi alcalina può essere adoperata anche per defosforilare i terminali 5′ del DNA o dell′RNA per prevenire l′auto-legatura. Il DNA e l′RNA possono anche essere marcati con il fosfato radiomarcato (tramite la T4 polinucleotide chinasi) dopo la defosforilazione con la fosfatasi alcalina.
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.
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.
Azioni biochim/fisiol
L′enzima ha un′ampia specificità per gli esteri fosforici di alcoli, ammine, pirofosfato e fenoli. Viene utilizzato abitualmente per defosforilare le proteine e gli acidi nucleici.
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 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.
Proprietà fisiche
La fosfatasi alcalina intestinale bovina è una glicoproteina dimerica derivata dalla membrana. Ne esistono almeno tre isoforme, che generalmente possiedono due glicani N-legati e uno o più glicani O-legati per monomero.
La fosfatasi alcalina intestinale bovina è una glicoproteina dimerica derivata dalla membrana. Ne esistono almeno tre isoforme, che generalmente possiedono due glicani N-legati e uno o più glicani O-legati per monomero.2 L′enzima richiede zinco e ioni bivalenti di magnesio o calcio per svolgere l′attività.
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.
Definizione di unità
Un′unità DEA idrolizza 1 μmole di 4-nitrofenil fosfato in un minuto a pH 9,8 e a 37 °C (un′unità di glicina è equivalente a circa 3 unità DEA).
Stato fisico
Solution in 3.0 M NaCl containing 5 mM MgCl2, 0.2 mM ZnCl2, and 30 mM triethanolamine, pH 7.6
Nota sulla preparazione
Affinity purified
Risultati analitici
Le dimensioni delle confezioni sono basate su unità DEA.
Protein determined by biuret.
Inibitore
Avvertenze
Danger
Indicazioni di pericolo
Consigli di prudenza
Classi di pericolo
Resp. Sens. 1
Codice della classe di stoccaggio
10 - Combustible liquids
Classe di pericolosità dell'acqua (WGK)
WGK 2
Punto d’infiammabilità (°F)
Not applicable
Punto d’infiammabilità (°C)
Not applicable
Dispositivi di protezione individuale
Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)
Certificati d'analisi (COA)
Cerca il Certificati d'analisi (COA) digitando il numero di lotto/batch corrispondente. I numeri di lotto o di batch sono stampati sull'etichetta dei prodotti dopo la parola ‘Lotto’ o ‘Batch’.
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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
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Protocolli
Enzymatic Assay of Alkaline Phosphatase, Diethanolamine Assay (EC 3. 1. 3. 1)
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