P5521

Phosphatase, Alkaline from bovine intestinal mucosa

≥2,000 DEA units/mg protein

• 동의어(들): Alkaline phosphatase, Orthophosphoric-monoester phosphohydrolase (alkaline optimum)
• CAS Number: 9001-78-9
• 효소 위원회 번호: 3.1.3.1 (BRENDA, IUBMB)
• MDL number: MFCD00131849
• UNSPSC 코드: 12352204
• NACRES: NA.54

속성

• 양식: aqueous solution
• Quality Level: 200
• 특이 활성도: ≥2,000 DEA units/mg protein
• 분자량: dimer ~160 kDa
• 농도: ≥1.0 mg/mL
• 저장 온도: 2-8°C

설명

일반 설명

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]

애플리케이션

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.

The enzyme from Sigma has been used in the assay of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate.[9] It has also been used for alkaline phosphatase assays using a Nafion^™-modified electrode.[10]

생화학적/생리학적 작용

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.

물리적 특성

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.

단위 정의

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)

물리적 형태

solution in 3.2 M ammonium sulfate, 1 mM MgCl₂ and 0.1 mM ZnCl₂, pH 7.0

제조 메모

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]

분석 메모

Package sizes are based on DEA units

Protein determined by biuret.

법적 정보

Nafion is a trademark of The Chemours Company FC, LLC

안전성 정보

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 1
• Flash Point (°F): Not applicable
• Flash Point (°C): Not applicable
• 개인 보호 장비: Eyeshields, Gloves, type N95 (US)