{"product_id":"alkaline-phosphatase","title":"Alkaline Phosphatase","description":"\n\u003ch2\u003eDescription\u003c\/h2\u003e\n\n\u003cp\u003eAlkaline phosphatases are ubiquitous enzymes, present in all species from bacteria to humans. In humans, four loci encode the different alkaline phosphatase isoenzymes.\u003c\/p\u003e\n\n\u003cp\u003eFor laboratories that require sensitive and reproducible alkaline phosphatase determination, the key practical consideration is the reaction behavior of the enzyme. In vitro, at neutral pH, alkaline phosphatase behaves as a \u003cstrong\u003ephosphotransferase\u003c\/strong\u003e rather than a hydrolase. Consequently, the reaction rate — and therefore the sensitivity of alkaline phosphatase determination — is considerably increased by using \u003cstrong\u003ephosphate-accepting molecules\u003c\/strong\u003e, such as diethanolamine, triethanolamine, Tris, and 2-amino-2-methyl-1-propanol.\u003c\/p\u003e\n\n\u003cp\u003eThis phosphate-acceptor requirement is the basis for the sensitivity of alkaline phosphatase determination, and it should be taken into account whenever setting up a dephosphorylation or conjugation reaction with this enzyme.\u003c\/p\u003e\n\n\u003ch2\u003eSpecification\u003c\/h2\u003e\n\n\u003cdiv style=\"overflow-x:auto;-webkit-overflow-scrolling:touch;\"\u003e\n  \u003ctable style=\"border-collapse:collapse;width:100%;min-width:560px;\"\u003e\n    \u003cthead\u003e\n      \u003ctr\u003e\n        \u003cth style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;font-weight:600;white-space:nowrap;\"\u003eProperty\u003c\/th\u003e\n        \u003cth style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;font-weight:600;white-space:nowrap;\"\u003eValue\u003c\/th\u003e\n      \u003c\/tr\u003e\n    \u003c\/thead\u003e\n    \u003ctbody\u003e\n      \u003ctr\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eSource\u003c\/td\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eCalf intestine\u003c\/td\u003e\n      \u003c\/tr\u003e\n      \u003ctr\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eGrade\u003c\/td\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eConjugation Grade\u003c\/td\u003e\n      \u003c\/tr\u003e\n      \u003ctr\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eForm\u003c\/td\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eAmmonium sulfate suspension\u003c\/td\u003e\n      \u003c\/tr\u003e\n      \u003ctr\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eSolubility\u003c\/td\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eDistilled water or diluted buffer\u003c\/td\u003e\n      \u003c\/tr\u003e\n      \u003ctr\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eStability\u003c\/td\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eStore at 4 °C (39 °F); do not freeze\u003c\/td\u003e\n      \u003c\/tr\u003e\n      \u003ctr\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eSpecific activity\u003c\/td\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003e~3,000–6,000 U\/mg protein\u003c\/td\u003e\n      \u003c\/tr\u003e\n      \u003ctr\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eCatalog No.\u003c\/td\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eBF-1404500\u003c\/td\u003e\n      \u003c\/tr\u003e\n      \u003ctr\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003ePrice\u003c\/td\u003e\n        \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003e$10.00\/mg\u003c\/td\u003e\n      \u003c\/tr\u003e\n    \u003c\/tbody\u003e\n  \u003c\/table\u003e\n\u003c\/div\u003e\n\n\u003cp\u003e\u003cstrong\u003eUnit definition:\u003c\/strong\u003e One unit (U) is the amount of enzyme that releases 1 μmol of p-nitrophenol per minute from p-nitrophenyl phosphate at 37 °C in diethanolamine buffer, pH 9.8.\u003c\/p\u003e\n\n\u003ch2\u003eFeatures\u003c\/h2\u003e\n\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eConjugation Grade\u003c\/strong\u003e — suitable for conjugation applications.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eHigh specific activity\u003c\/strong\u003e — approximately 3,000–6,000 U\/mg protein.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003ePhosphotransferase activity at neutral pH\u003c\/strong\u003e — the reaction rate is considerably increased by phosphate-accepting molecules (diethanolamine, triethanolamine, Tris, 2-amino-2-methyl-1-propanol).\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eAmmonium sulfate suspension\u003c\/strong\u003e — stable storage format.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSoluble\u003c\/strong\u003e in distilled water or diluted buffer.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eStable at 4 °C (39 °F)\u003c\/strong\u003e — do not freeze.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eApplication\u003c\/h2\u003e\n\n\u003cp\u003eAlkaline Phosphatase, Calf Intestine (Conjugation Grade) is intended for \u003cstrong\u003econjugation\u003c\/strong\u003e applications, in which the enzyme is coupled to antibodies and proteins. The enzyme catalyzes the \u003cstrong\u003edephosphorylation\u003c\/strong\u003e of phosphate groups and, at neutral pH, behaves as a phosphotransferase.\u003c\/p\u003e\n\n\u003ch2\u003eFAQ\u003c\/h2\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eWhat is the source and grade of this alkaline phosphatase?\u003c\/summary\u003e\n  \u003cp\u003eThis alkaline phosphatase is obtained from calf intestine and is supplied at Conjugation Grade.\u003c\/p\u003e\n\u003c\/details\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eWhat is the specific activity of this alkaline phosphatase?\u003c\/summary\u003e\n  \u003cp\u003eThe specific activity is approximately 3,000–6,000 U\/mg protein.\u003c\/p\u003e\n\u003c\/details\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eWhat is the unit definition of alkaline phosphatase?\u003c\/summary\u003e\n  \u003cp\u003eOne unit (U) is the amount of enzyme that releases 1 μmol of p-nitrophenol per minute from p-nitrophenyl phosphate at 37 °C in diethanolamine buffer, pH 9.8.\u003c\/p\u003e\n\u003c\/details\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eHow is alkaline phosphatase activity measured?\u003c\/summary\u003e\n  \u003cp\u003eAlkaline phosphatase activity is measured by following the increase in absorbance at 405 nm, which corresponds to the release of p-nitrophenol from p-nitrophenyl phosphate.\u003c\/p\u003e\n\u003c\/details\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eWhat reagents are required for the activity assay?\u003c\/summary\u003e\n  \u003cdiv style=\"overflow-x:auto;-webkit-overflow-scrolling:touch;\"\u003e\n    \u003ctable style=\"border-collapse:collapse;width:100%;min-width:560px;\"\u003e\n      \u003cthead\u003e\n        \u003ctr\u003e\n          \u003cth style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;font-weight:600;white-space:nowrap;\"\u003eReagent\u003c\/th\u003e\n          \u003cth style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;font-weight:600;white-space:nowrap;\"\u003ePreparation\u003c\/th\u003e\n        \u003c\/tr\u003e\n      \u003c\/thead\u003e\n      \u003ctbody\u003e\n        \u003ctr\u003e\n          \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eDiethanolamine buffer (1 mol\/L, pH 9.8; 0.5 mmol\/L MgCl\u003csub\u003e2\u003c\/sub\u003e)\u003c\/td\u003e\n          \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003e9.7 mL diethanolamine (99%) + distilled water; add 0.05 mL MgCl\u003csub\u003e2\u003c\/sub\u003e solution; adjust to pH 9.8 (37 °C) with HCl (≥2 mol\/L); make up to 100 mL.\u003c\/td\u003e\n        \u003c\/tr\u003e\n        \u003ctr\u003e\n          \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eMgCl\u003csub\u003e2\u003c\/sub\u003e solution (1 mol\/L)\u003c\/td\u003e\n          \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003e20.3 g MgCl\u003csub\u003e2\u003c\/sub\u003e·6H\u003csub\u003e2\u003c\/sub\u003eO dissolved in 100 mL distilled water.\u003c\/td\u003e\n        \u003c\/tr\u003e\n        \u003ctr\u003e\n          \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003e4-Nitrophenyl phosphate solution (0.67 mol\/L)\u003c\/td\u003e\n          \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003e250 mg 4-nitrophenyl phosphate sodium salt + 1.0 mL distilled water.\u003c\/td\u003e\n        \u003c\/tr\u003e\n        \u003ctr\u003e\n          \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eDilution buffer (0.1 mol\/L, pH 7.6)\u003c\/td\u003e\n          \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003e1.86 g TEA·HCl in distilled water; add 0.1 mL MgCl\u003csub\u003e2\u003c\/sub\u003e solution + 0.1 mL ZnCl\u003csub\u003e2\u003c\/sub\u003e (0.1 mol\/L, freshly prepared); adjust to pH 7.6 with NaOH (1 mol\/L); make up to 100 mL.\u003c\/td\u003e\n        \u003c\/tr\u003e\n        \u003ctr\u003e\n          \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eSample solution\u003c\/td\u003e\n          \u003ctd style=\"border:1px solid #ccc;padding:8px 10px;text-align:left;vertical-align:top;\"\u003eDilute with dilution buffer to 0.05–0.06 U\/mL; stand for 15–20 min at room temperature before assay.\u003c\/td\u003e\n        \u003c\/tr\u003e\n      \u003c\/tbody\u003e\n    \u003c\/table\u003e\n  \u003c\/div\u003e\n\u003c\/details\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eWhat is the assay procedure for alkaline phosphatase?\u003c\/summary\u003e\n  \u003col\u003e\n    \u003cli\u003eSet the spectrophotometer to 405 nm and 37 °C.\u003c\/li\u003e\n    \u003cli\u003eInto a cuvette, add 2.90 mL diethanolamine buffer and 0.05 mL 4-nitrophenyl phosphate solution.\u003c\/li\u003e\n    \u003cli\u003eIncubate at 37 °C for 6–8 minutes (temperature equilibration and blank rate).\u003c\/li\u003e\n    \u003cli\u003eAdd 0.1 mL diluted enzyme solution, mix, and record the absorbance increase at 405 nm for 5 minutes.\u003c\/li\u003e\n    \u003cli\u003eUse the linear portion of the curve to calculate ΔE\u003csub\u003e405\u003c\/sub\u003e\/min.\u003c\/li\u003e\n  \u003c\/ol\u003e\n\u003c\/details\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eHow is the activity of alkaline phosphatase calculated?\u003c\/summary\u003e\n  \u003cp\u003eActivity (U\/mg) = (ΔE\u003csub\u003e405\u003c\/sub\u003e\/min × total volume × enzyme dilution) \/ (18.5 × enzyme volume × mg enzyme\/mL).\u003c\/p\u003e\n  \u003cp\u003eHere, 18.5 is the millimolar extinction coefficient of p-nitrophenol at 405 nm.\u003c\/p\u003e\n\u003c\/details\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eHow should this alkaline phosphatase be stored?\u003c\/summary\u003e\n  \u003cp\u003eStore at 4 °C (39 °F). Do not freeze.\u003c\/p\u003e\n\u003c\/details\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eIn what form is this alkaline phosphatase supplied?\u003c\/summary\u003e\n  \u003cp\u003eThe product is supplied as an ammonium sulfate suspension.\u003c\/p\u003e\n\u003c\/details\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eWhat is the solubility of this alkaline phosphatase?\u003c\/summary\u003e\n  \u003cp\u003eThe enzyme is soluble in distilled water or diluted buffer.\u003c\/p\u003e\n\u003c\/details\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eWhy does alkaline phosphatase behave as a phosphotransferase at neutral pH?\u003c\/summary\u003e\n  \u003cp\u003eIn vitro, at neutral pH, alkaline phosphatase behaves as a phosphotransferase rather than a hydrolase. This behavior means that the reaction rate depends on the availability of a phosphate-accepting molecule.\u003c\/p\u003e\n\u003c\/details\u003e\n\n\u003cdetails\u003e\n  \u003csummary style=\"cursor:pointer;font-weight:600;\"\u003eWhich phosphate-accepting molecules increase the reaction rate?\u003c\/summary\u003e\n  \u003cp\u003ePhosphate-accepting molecules such as diethanolamine, triethanolamine, Tris, and 2-amino-2-methyl-1-propanol considerably increase the reaction rate. This is the basis for the sensitivity of alkaline phosphatase determination.\u003c\/p\u003e\n\u003c\/details\u003e\n","brand":"calzyme","offers":[{"title":"500 mg","offer_id":52982411198645,"sku":"BF-1404500","price":0.0,"currency_code":"USD","in_stock":true},{"title":"1 g","offer_id":52982411231413,"sku":"BF-1404501","price":0.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0521\/5312\/2997\/files\/alkaline-phosphatase-BF-1404500-1500x1500.jpg?v=1787042827","url":"https:\/\/biofargo.com\/products\/alkaline-phosphatase","provider":"Biofargo","version":"1.0","type":"link"}