{"product_id":"lipase-enzyme-activity-tbp0077","title":"Lipase, Enzyme Activity","description":"\u003cp\u003e\u003cspan style=\"font-size: 18px\"\u003e\u003cstrong\u003e\u003cu\u003eProduct Details\u003c\/u\u003e\u003c\/strong\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eForm:\u003c\/u\u003e Freeze-dried\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eSolubility:\u003c\/u\u003e Soluble in water and dilute buffer\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eStability:\u003c\/u\u003e -20° C; -4° F\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eActivity:\u003c\/u\u003e 20,000-50,000\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eProtein:\u003c\/u\u003e 90%\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size: 18px\"\u003e\u003cstrong\u003e\u003cu\u003eUnit Definition\u003c\/u\u003e\u003c\/strong\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003eThe amount of enzyme causing the liberation of one micromole of glycerol per minute at 37°C and pH 7.0.\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size: 18px\"\u003e\u003cstrong\u003e\u003cu\u003eAssay Method\u003c\/u\u003e\u003c\/strong\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003eThe assay is based on a 4-step enzyme-coupled reaction. In the final step of the sequence, a dye is formed which can be measured spectrophotometrically at 490 nm. The increase in absorbance at 490 nm is directly proportional to lipase activity.\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size: 18px\"\u003e\u003cstrong\u003e\u003cu\u003eApplications\u003c\/u\u003e\u003c\/strong\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003eLipase (EC 3.1.1.3) hydrolyses emulsified triglycerides of the long chain fatty acids. Lipase is active at the interface between the oil drops and the aqueous phase. In normal serum the concentration of lipase is low. In acute pancreatitis and in pancreatic carcinoma a rise in serum lipase activity occurs, with a mean increase being 50 times that of normal values. A rise in the serum lipase content is also found in acute and chronic renal diseases.\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size: 18px\"\u003e\u003cstrong\u003e\u003cu\u003eReagents\u003c\/u\u003e\u003c\/strong\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003col\u003e\n\u003cli\u003e0.1 M Potassium phosphate buffer, pH 7.0.\u003c\/li\u003e\n\u003cli\u003e0.05 M 4-Aminoantipyrine-HCl (12 mg\/ml) in distilled H2O.\u003c\/li\u003e\n\u003cli\u003e0.05 M 1,7-Dihydroxynaphthalene (8 mg\/ml absolute ethanol).\u003c\/li\u003e\n\u003cli\u003e0.05 M Magnesium sulfate (6.02 g\/1000 ml) in distilled H2O.\u003c\/li\u003e\n\u003cli\u003e0.05 M ATP (30.3 mg\/ml) in distilled H2O.\u003c\/li\u003e\n\u003cli\u003ePeroxidase solution (588 U\/ml). Dilute in 0.05 M phosphate buffer, pH 7.0 to a final concentration of 588 U\/ml.\u003c\/li\u003e\n\u003cli\u003eGlycerokinase solution (36 U\/ml). Dilute in 0.05 M phosphate buffer, pH 7.0 to a final concentration of 36 U\/ml.\u003c\/li\u003e\n\u003cli\u003ea-Glycerophosphate Oxidase solution (700 U\/ml). Dilute in distilled H2O to a final concentration of 700 U\/ml.\u003c\/li\u003e\n\u003cli\u003eTriolein emulsion. Mix 300 mg triolein\/10.6 g Triton X-100 and heat until a single phase appears. Add 90 ml of distilled H2O and mix.\u003c\/li\u003e\n\u003cli\u003eReaction mixture. Mix together 0.128 ml 4-Aminoantipyrine, 0.064 ml 1,7-Dihydroxynaphthalene, 0.128 ml peroxidase, 12.4 ml 0.1 M phosphate buffer, pH 7.0, 0.150 ml glycerokinase, 0.32 ml a-glycerophosphate oxidase, 0.375 ml triolein emulsion, 0.375 ml magnesium sulfate and 0.375 ml ATP. Equilibrate at 4¡C for one hour before use. Stable for 8 hours at 4¡C.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"text-decoration: underline\"\u003e\u003cstrong\u003e\u003cspan style=\"font-size: 18px\"\u003eProcedure\u003c\/span\u003e\u003c\/strong\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003col\u003e\n\u003cli\u003eSet spectrophotometer (equipped with strip chart recorder and temperature control) at 490 nm and 37°C.\u003c\/li\u003e\n\u003cli\u003eInto a cuvette pipette 2.95 ml of reaction mixture and equilibrate at 37°C for 10 min.\u003c\/li\u003e\n\u003cli\u003eDissolve 10 mg of lyophilized lipase in 10 ml of distilled H2O and dilute 1:100 with distilled H2O.\u003c\/li\u003e\n\u003cli\u003eAdd 0.05 ml of the lipase solution to the equilibrated cuvette and mix.\u003c\/li\u003e\n\u003cli\u003eRecord the change in absorbance at 490 nm for 10 min.\u003c\/li\u003e\n\u003cli\u003eMeasure the slope after 5 min. If (delta)A\/min. is greater than 0.015, a larger dilution is required. If the (delta)A\/min. is less than 0.005, a smaller dilution is required.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003c!--TBP0077-100KU TBP0077-500KU--\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size: 13px; font-weight: 700; text-transform: uppercase;\"\u003e\u003cstrong\u003eResearch use only.\u003c\/strong\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003ch2\u003eDocuments\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/tribioscience.com\/wp-content\/uploads\/2024\/03\/TBP0077-Lipase-1.pdf\"\u003eDatasheet (PDF)\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/tribioscience.com\/wp-content\/uploads\/2024\/03\/TBP0077_Lipase-Safety-Data-Sheet-1.pdf\"\u003eMSDS (PDF)\u003c\/a\u003e\u003c\/li\u003e\n\u003c\/ul\u003e","brand":"TriBioScience","offers":[{"title":"100 KU","offer_id":45986499690575,"sku":"TBP0077-100KU","price":0.0,"currency_code":"ILS","in_stock":true},{"title":"500 KU","offer_id":45986499723343,"sku":"TBP0077-500KU","price":0.0,"currency_code":"ILS","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0712\/0748\/5519\/files\/TBP0077.png?v=1782812660","url":"https:\/\/www.danyel.co.il\/products\/lipase-enzyme-activity-tbp0077","provider":"danyel biotech","version":"1.0","type":"link"}