{"product_id":"apo-d-amino-acid-oxidase-enzyme-activity-tbp0056","title":"Apo D-Amino Acid Oxidase, 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 powder\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 -20° C (-4° F)\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eActivity:\u003c\/u\u003e 25-30 U\/mg protein\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eProtein:\u003c\/u\u003e 90%\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eContaminants:\u003c\/u\u003e Free of FAD and Alkaline Phosphatase\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 that will deaminate by oxidation one micromole of D-alanine to pyruvate per minute at pH 8.3, at 37°C in the presence of catalase.\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 the method described by Bergmeyer. The decrease in the absorbance at 340 nm, due to the oxidation of NADH, is a measure of D-amino acid oxidase 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\u003eThe D isomers of alanine, methionine, valine, isoleucine, phenylalanine and proline serve as good substrates while the L isomers do not react at all. The enzyme is a flavoprotein. D-amino acid oxidase from porcine kidney has been extensively studied. It has a monomeric molecular weight of 38,000-39,000.\u003c\/p\u003e\n\u003cp\u003eD-Amino acid oxidase (EC 1.4.3.3) has several possible applications such as the determination of D-amino acids, the separation of natural L- amino acid isomers from a racemic mixture and in the preparation of keto acids. The usefulness and application of D-amino acid oxidase can be significantly increased if it is available in an immobilized form.\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\u003e0.2 M Tris-HCl buffer, pH 8.3.\u003c\/li\u003e\n\u003cli\u003e0.02 M D-Alanine (17.8 mg\/ml) in buffer.\u003c\/li\u003e\n\u003cli\u003e0.008 M NADH disodium salt (5 mg\/ml) in buffer.\u003c\/li\u003e\n\u003cli\u003eCatalase (200 U\/ml) in buffer. Prepare fresh.\u003c\/li\u003e\n\u003cli\u003eLactate dehydrogenase (LDH) (200 U\/ml) in buffer. Prepare fresh.\u003c\/li\u003e\n\u003cli\u003eFAD (Prepare1 mg\/ml solution)\u003c\/li\u003e\n\u003cli\u003eD-Amino acid oxidase solution. Dilute in buffer to give a concentration of 0.1-0.5 U\/ml. Must be prepared fresh prior to assay.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cspan style=\"text-decoration: underline\"\u003e\u003cspan style=\"font-size: 18px\"\u003eProcedure\u003c\/span\u003e\u003c\/span\u003e\u003c\/strong\u003e\u003c\/p\u003e\n\u003col\u003e\n\u003cli\u003eSet spectrophotometer (equipped with a strip chart recorder and temperature control) at 340 nm and 37°C.\u003c\/li\u003e\n\u003cli\u003eBubble oxygen through the buffer for 5-10 min. to saturate it with oxygen.\u003c\/li\u003e\n\u003cli\u003eIn a cuvette, pipette the following reagents in the amounts indicated:\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"padding-left: 40px\"\u003eTris buffer (oxygenated) 2.00 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eD-Alanine 0.50 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eNADH 0.10 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eCatalase 0.10 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eLDH 0.10 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eFAD 0.10 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eIncubate in spectrophotometer at 37°C for 5 min. to attain temperature equilibration. Record absorbance at 340 nm (blank).\u003c\/p\u003e\n\u003col start=\"4\"\u003e\n\u003cli\u003eInitiate the reaction by adding 0.1 ml D-amino acid oxidase (enzyme) to the cuvette. Follow the reaction by recording the decrease in the absorbance at 340 nm for 5-8 min.\u003c\/li\u003e\n\u003cli\u003eCalculate E340nm\/min\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e\u003c!--TBP0056-100U TBP0056-500U--\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\/TBP0056-Apo-D-Amino-Acid-Oxidase-1.pdf\"\u003eDatasheet (PDF)\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/tribioscience.com\/wp-content\/uploads\/2024\/03\/TBP0056_Apo-D-Amino-Acid-Oxidase-Safety-Data-Sheet-1.pdf\"\u003eMSDS (PDF)\u003c\/a\u003e\u003c\/li\u003e\n\u003c\/ul\u003e","brand":"TriBioScience","offers":[{"title":"100 U","offer_id":45986497396815,"sku":"TBP0056-100U","price":0.0,"currency_code":"ILS","in_stock":true},{"title":"500 U","offer_id":45986497429583,"sku":"TBP0056-500U","price":0.0,"currency_code":"ILS","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0712\/0748\/5519\/files\/TBP0056.png?v=1782812619","url":"https:\/\/www.danyel.co.il\/products\/apo-d-amino-acid-oxidase-enzyme-activity-tbp0056","provider":"danyel biotech","version":"1.0","type":"link"}