Uricase (1711) - Sekisui Diagnostics

Page 1 of 3UricaseCatalogue No. 70-1711-01Origin:Candida utilisSpecifications:Appearance:White to off white free flowing powderSpecific Activity: > 6 U/mg protein at 25°CCharacteristics:Molecular weight:120k Da (Gel Permeation Chromatography)32-35 kDa (SDS-PAGE)Isoelectric point: 5.6Optimum pH: 7.5 – 9.5 See Fig.1pH Stability: 6.5 – 11 (5°C, 1 week) See Fig.2Optimum Temperature: Above 35°C See Fig.3Thermal Stability: Below 50°C (pH 8.5, 10 min) See Fig.4Lyophilised Stability: Stable for 12 months desiccated at -20°CFig. 1 Effect of various buffer solutions on Uricase Activity0.250.2∆ A 293 nm/min0.150.1K-phosphate bufferTris-HCL bufferGly-KOH bufferGTA bufferTEA HCL-KOH bufferBorate buffer0.0505 6 7 8 9 10 11pH(80mM buffer, 0.125mM Urate)AP0198.1.0Intended for use in the manufacture of IVDs

Page 2 of 3Uricase(Catalogue No. 1711)Fig. 2 pH Stability0.20.15∆ A 293 nm/min0.1K-citrate bufferK-phosphate bufferTris-HCL bufferGly-KOH bufferGTA bufferTEA-HCL-KOH buffer0.0503 4 5 6 7 8 9 10 11 12pHStored at 5°C for 1 week in 100mM buffer(80mM Borate buffer (pH 8.5), 0.125mM Urate)Fig. 3 Effect of temperature on Uricase activity with borate buffer250200Relative activity15010050024 26 28 30 32 34 36 38Temperature (degrees centigrade)(50mM Borate buffer (pH 8.5), 0.125mM Urate)AP0198.1.0Intended for use in the manufacture of IVDs

Page 3 of 3Uricase(Catalogue No. 1711)Fig. 4 Thermal Stability0.2∆ A 293 nm/min0.100 10 20 30 40 50 60 70 80 90Temperature (degree centigrade)(10 minute incubation in 50mM borate buffer, pH 8.5, 0.1mM Uric acid)Assay Principle:Uricase catalyses the following reaction:UricaseUric Acid + 2H 2 O 2 + O 2 Allantoin + H 2 O 2 + CO 2As uric acid degrades there is a decrease in absorbance which can be measuredspectrophotometrically at 290nm.Unit Definition:One unit of activity is defined as the amount of enzyme that will transform 1.0 micromole ofsubstrate per minute at 25ºC under standard assay method conditions.(See Analytical Method for full details)International50 Gibson DriveKings Hill, West MallingKent, ME19 4AF, UKPhone: +44 (0) 1732 220022Fax: +44 (0) 1732 220024/5The Americas31 New York AvenueFramingham, MA01701-9322Phone: 800 332 1042Fax: 800 762 6311AP0198.1.0Intended for use in the manufacture of IVDs