{"product_id":"leucine-aminopeptidase-enzyme-activity-tbp0076","title":"Leucine Aminopeptidase, 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 distilled water or 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 U\/mg protein\u003c\/p\u003e\n\u003cp\u003e\u003cu\u003eProtein:\u003c\/u\u003e 90%\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\u003eThat amount of enzyme which catalyzes the hydrolysis of one micromole L-leucinamide per minute at 25°C at pH 8.5.\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 method is based on the one described by Binkley and Torres. The rate of decrease in absorbance at 238 nm, resulting from the hydrolysis of L-leucinamide, is proportional to the catalytic activity of the enzyme.\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\u003eLeucine aminopeptidase (LAP) is a proteolytic enzyme which hydrolyzes the peptide bond adjacent to a free amino group. It is called leucine aminopeptidase because it rapidly catalyzes the hydrolysis of leucine containing peptides. However, it also catalyzes the hydrolytic release of other amino acids located at the N-terminal end of various peptides and proteins. The enzyme from porcine kidney has been extensively studied. It has a molecular weight of 255,000 and it consists of four subunits each having one atom of zinc.\u003c\/p\u003e\n\u003cp\u003eDetermination of microsomal leucine amino peptidase activity in serum is of clinical significance, since serum LAP levels are elevated in obstructive jaundice, liver cirrhosis, liver carcinoma and also during the later part of pregnancy. Leucine amino-peptidase is also extensively used in determination of the amino acid sequence of proteins and peptides.\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\u003e0.5 M Tris-HCl buffer, pH 8.5.\u003c\/li\u003e\n\u003cli\u003e0.025 M Maganese chloride.\u003c\/li\u003e\n\u003cli\u003e0.0625 M Magnesium chloride.\u003c\/li\u003e\n\u003cli\u003e0.06 M L-leucinamide-HCl (11 mg\/ml) in buffer.\u003c\/li\u003e\n\u003cli\u003eLeucine aminopeptidase (enzyme) solution. Dissolve in buffer to a concentration of 1000-2000 U\/ml.\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 238 nm and 25°C.\u003c\/li\u003e\n\u003cli\u003eEnzyme treatment. In a test tube pipette the following:\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"padding-left: 40px\"\u003eManganese chloride 0.4 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eTris buffer 2.5 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eEnzyme (LAP) 0.1 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eIncubate the mixture at 40°C for about 60 minutes. An aliquot from this treated enzyme mixture will be used in the assay system.\u003c\/p\u003e\n\u003col start=\"3\"\u003e\n\u003cli\u003eIn a quartz cuvette, place the following in the amounts indicated:\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp style=\"padding-left: 40px\"\u003eTris buffer 0.20 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eMagnesium chloride 0.20 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003eL-leucinamide-HCl 2.50 ml\u003c\/p\u003e\n\u003cp style=\"padding-left: 40px\"\u003ePlace cuvette in spectrophotometer at 25°C for 5 minutes. Record absorbance at 238 nm (blank).\u003c\/p\u003e\n\u003col start=\"4\"\u003e\n\u003cli\u003eTo initiate the reaction, add 0.10 ml of treated enzyme (step 2) to the cuvette. Follow the reaction by recording the decrease in absorbance at 238 nm for 5-8 minutes.\u003c\/li\u003e\n\u003cli\u003eCalculate (delta)E238nm\/min\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003c!--TBP0076-100U TBP0076-500U--\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\/TBP0076-Leucine-Aminopeptidase-1.pdf\"\u003eDatasheet (PDF)\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/tribioscience.com\/wp-content\/uploads\/2024\/03\/TBP0076_Leucine-Aminopeptidase-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":45986499788879,"sku":"TBP0076-100U","price":0.0,"currency_code":"ILS","in_stock":true},{"title":"500 U","offer_id":45986499821647,"sku":"TBP0076-500U","price":0.0,"currency_code":"ILS","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0712\/0748\/5519\/files\/TBP0076.png?v=1782812660","url":"https:\/\/www.danyel.co.il\/products\/leucine-aminopeptidase-enzyme-activity-tbp0076","provider":"danyel biotech","version":"1.0","type":"link"}