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外観
液体
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解説
ALPともいう。有機化合物であるリン酸エステルを加水分解する酵素。高等動物の場合には，臓器特異性をもった ALPのアイソザイム (同位酵素) が存在する。生理作用は不明であるが，血清中にはアルカリ性と酸性の2種のホスファターゼがあり，このうちアルカリ性のものは肝臓や胆道，骨などに由来し，これらの臓器に疾患があると血清中に増量する。この性質を利用して，肝機能検査などに広く用いられる。
ブリタニカ国際大百科事典 小項目事典 ブリタニカ
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用途
EIA用標識酵素, 化学発光法によるりん脂質の臨床分析への応用。
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用途
EIA用標準酵素
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生物学のな役割
アルカリホスファターゼ［EC3.1.3.1］．アルカリ性ホスファターゼともいう．アルカリ性領域に至適pHをもつホスホモノエステラーゼ．亜鉛酵素で，特異性の広いホスファターゼ．広く生物の膜画分に存在する．妊娠後期や骨症のときなどに血漿中の濃度が上昇する．
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使用上の注意
安定性安定pH:6～9
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化学的特性
solid
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使用
Alkaline Phosphatase is commonly used to remove the 5′-terminal phosphate from nucleic acids.
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一般的な説明
Alkaline phosphatase from bovine intestinal mucosa is both a phosphomonoesterase and a pyrophosphatase.
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純化方法
The E.coli supernatant in sucrose (20%, 33mM) in Tris-HCl pH 8.0 is purified through a DEAE-cellulose column and recrystallised. To the column eluates in 0.125M NaCl is added MgCl2 (to 0.01M) and brought to 50% saturation in (NH4)2SO4 by adding the solid (0.20g/mL). The mixture is centrifuged to remove bubbles and is adjusted to pH 8.0 (with 2N NaOH). Saturated (NH4)2SO4 at pH 8.0 is added dropwise until the solution becomes faintly turbid (~61% saturation). It is set aside at ~25o for 1hour (turbidity will increase). The mixture is placed in an ice bath for several minutes when turbidity disappears and a clear solution is obtained. It is then placed in a large ice bath at 0o (~5L) and allowed to warm slowly to room temperature in a dark room whereby crystals are formed appearing as a silky sheen. The crystals are collected by centrifugation at 25o if necessary. The crystalline solutions are stable at ~25o for many months. They can be stored at 0o, but are unstable when frozen. Cysteine at 10-3M and thioglycolic acid at 10-4M are inhibitory. This is reversed on addition of Zn2+ ions. Many organic phosphates are good substrates for this phosphatase. [Molamy & Horecker Methods Enzymol 9 639 1966, Torriani et al. Methods Enzymol 12b 212 1968, Engstrom Biochim Biophys Acta 92 71 1964.] Alkaline phosphatase from rat osteosarcoma has been purified by Me2CO precipitation and chromatography on DEAE-cellulose, Sephacryl S-200, and hydroxylapatite. [Nair et al. Arch Biochem Biophys 254 18 1987.] Phosphoproteins (various). These are purified by adsorbing onto an iminodiacetic acid substituted agarose column to which are bound ferric ions. This chelate complex acts as a selective immobilised metal affinity adsorbent for phosphoproteins. [Muszyfiska et al. Biochemistry 25 6850 1986.]
