{"product_id":"glutamic-oxaloacetic-transaminase-enzyme-activity-tbp0069","title":"Glutamic Oxaloacetic Transaminase, 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 Ammonium Sulfate\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eSolubility:\u003c\/u\u003e Distilled water or dilute buffer\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eStability:\u003c\/u\u003e Store at 4° C (39° F); Do not freeze\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eActivity:\u003c\/u\u003e 4000 U\/ml protein\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eProtein:\u003c\/u\u003e 10 mg\/ml\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eContaminants:\u003c\/u\u003e Glutamate dehydrogenase \u0026lt;0.01%;\u003c\/p\u003e\n\u003cp style=\"padding-left: 100px\"\u003eGlutamate pyruvate transaminase \u0026lt;0.01%;\u003c\/p\u003e\n\u003cp style=\"padding-left: 100px\"\u003eLactate dehydrogenase \u0026lt;0.01%;\u003c\/p\u003e\n\u003cp style=\"padding-left: 100px\"\u003eMalate dehydrogenase \u0026lt;0.01%;\u003c\/p\u003e\n\u003cp style=\"padding-left: 100px\"\u003eOxaloacetate decarboxylase \u0026lt;0.05%\u003c\/p\u003e\n\u003cp\u003e \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 which will catalyze the transamination of one micromole of L-aspartate per minute at 25°C and pH 7.5.\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 coupled reaction where the oxaloacetate formed is reduced to malate in the presence of NADH and malate dehydrogenase. The decrease in the absorbance at 340 nm caused by the oxidation of NADH is proportional to the catalytic activity of GOT.\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\u003e(L-Aspartate: a-Ketoglutarate amino transferase, EC 2.6.1.1) Glutamic oxaloacetic transaminase (GOT), also known as aspartate aminotransferase, catalyzes the following reaction:\u003c\/p\u003e\n\u003cp\u003eGOT L-Asp + a-Ketoglutarate ——\u0026gt; Oxaloacetate + L-Glu\u003c\/p\u003e\n\u003cp\u003eThe enzyme is widely distributed in plants and animals, but it occurs in concentrated form in mammalian heart and liver. GOT requires pyridoxal phosphate as a coenzyme for its activity. It exists as two isozyme forms, the mitochondrial form (M-GOT) and the cytosol form (S-GOT). GOT from porcine heart has a molecular weight in the range of 91,000-94,000. Serum GOT levels in healthy subjects are low, but the levels are significantly elevated in a number of clinical conditions such as acute and chronic hepatitis, obstructive jaundice, carcinoma of liver, myocardial infraction and muscular dystrophy. Therefore, determination of serum GOT level has great clinical and diagnostic significance. In addition, GOT has applications in coupled enzyme reactions for measurement of metabolite levels in biological fluids.\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 7.5.\u003c\/li\u003e\n\u003cli\u003e0.008 M NADH, disodium salt (5 mg\/ml) in buffer. Prepare fresh.\u003c\/li\u003e\n\u003cli\u003e0.3 M L-Aspartate-monopotassium salt (51.4 mg\/ml) in buffer.\u003c\/li\u003e\n\u003cli\u003e0.60 M a-Ketoglutarate (87 mg\/ml) in buffer.\u003c\/li\u003e\n\u003cli\u003eMalate dehydrogenase (MDH). Using phosphate buffer, prepare a solution to yield a concentration of 200-250 U\/ml. Prepare fresh prior to assay.\u003c\/li\u003e\n\u003cli\u003eGlutamate-oxaloacetate transaminase (GOT) enzyme solution. Using phosphate buffer, prepare a GOT solution with a concentration in the range of 0.2-0.4 U\/ml. Must be prepared fresh prior to the assay.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"font-size: 18px\"\u003e\u003cstrong\u003e\u003cu\u003eProcedure\u003c\/u\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 340 nm and 25°C.\u003c\/li\u003e\n\u003cli\u003eInto a cuvette pipette the following reagents in the amounts indicated:\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"padding-left: 40px\"\u003ePhosphate buffer 1.00 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eNADH 0.10 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eL-Aspartate 1.50 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003ea-ketoglutarate 0.20 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eMDH 0.10 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eMix and incubate in spectrophotometer at 25°C for 5 minutes.\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003col start=\"3\"\u003e\n\u003cli\u003eRecord absorbance at 340 nm (blank), if any.\u003c\/li\u003e\n\u003cli\u003eInitiate the reaction by adding 0.1 ml of enzyme (GOT) solution. Record absorbance at 340 nm for 5 minutes.\u003c\/li\u003e\n\u003cli\u003eCalculate (delta)E340 nm\/min\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003c!--TBP0069-1KU TBP0069-5KU--\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\/TBP0069-Glutamic-Oxaloacetic-Transaminase_datasheet.pdf\"\u003eDatasheet (PDF)\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/tribioscience.com\/wp-content\/uploads\/2024\/03\/TBP0069_Glutamic-Oxaloacetic-Transaminase-Safety-Data-Sheet-1.pdf\"\u003eMSDS (PDF)\u003c\/a\u003e\u003c\/li\u003e\n\u003c\/ul\u003e","brand":"TriBioScience","offers":[{"title":"1000 U","offer_id":45986498871375,"sku":"TBP0069-1000U","price":0.0,"currency_code":"ILS","in_stock":true},{"title":"5000 U","offer_id":45986498904143,"sku":"TBP0069-5000U","price":0.0,"currency_code":"ILS","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0712\/0748\/5519\/files\/TBP0069-5KU.jpg?v=1782812644","url":"https:\/\/www.danyel.co.il\/products\/glutamic-oxaloacetic-transaminase-enzyme-activity-tbp0069","provider":"danyel biotech","version":"1.0","type":"link"}