Certificate of Analysis
Contents
1. 9 gt Biochain www biochain com Tel 1 888 762 2568 Fax 1 510 783 5386 Email info biochain com User s Manual and Instructions Uric Acid Assay Kit Z5030021 DESCRIPTION Uric acid is the waste product produced from the degradation of purines In healthy human uric acid is filtered and removed from the blood by the kidneys and excreted into urine Because a number of kidney diseases are known to affect uric acid levels uric acid determination is thus important and useful in diagnosing and evaluating kidney diseases For example when uric acid is present in the blood at abnormally high levels it tends to crystallize in body joints resulting in gout a very painful inflammatory condition Increased levels of uric acid are also known to be associated with uremia leukemia and pneumonia Simple direct and automation ready procedures for measuring uric acid concentration in blood are becoming popular in Research and Drug Discovery Biochain s uric acid assay kit is designed to measure uric acid directly in serumwithout any pretreatment The improved method utilizes 2 4 6 tripy ridyl s triazine that forms a blue colored complex specifically with iron in the presence of uric acid The intensity of the color measured at 590nm is directly proportional to the uric acid concentration in the serum The optimized formulation substantially reduces interference by substances in the raw samples KEY FEATURES Sensitive and accurate Use 52. uL samples Linear detection range 0 22 mg dL 13uM to 30 mg dL 2380uM uric acid in 96 well plate assay Simple and high throughput The procedure involves addition of a single working reagent and incubation for 30 min Can be readily automated as a high throughput assay in 96 well plates for thousands of samples per day Improved reagent stability and versatility The optimized formulation has greatly enhanced reagent and signal stability Cuvet or 96 well plate assay Low interference in biological samples No pretreatments are needed Assays canbe directly performed on serum samples APPLICATIONS Direct Assays uric acid inserum plasma urine and other biolbgical samples Drug Discovery Pharmacology effects of drugs on uric acid metabolism KIT CONTENTS 250 tests in 96 well plates Reagent A 50 mL Reagent B 6 mL Reagent C 6mL Standard 1 mL 10mg dL uric acid Blank Control 1mL Storage conditions The kit is shipped at room temperature Store reagents at 4 C standard and blank control at 20 C Shelf life 12 months after receipt Precautions reagents are for research use only Normal precautions for laboratory reagents should be exercised while using the reagents Please refer to Material Safety Data Sheet for detailed information PROCEDURES Reagent Preparation shake Reagent C before use Prepare enough working reagent by mixing 10 volumes of Reagent A 1 volume Reagent B and 1 volume Reagent C Fresh reconstitut
3. 590nm reader Procedure using cuvette Cuvets for measuring optical density at 510 630nm Spectrophotometer for measuring absorbance at 590nm EXAMPLES Samples were assayed using the 96 well protocol The uric acid content mg dL was 1 3 0 1 n 4 for mice serum 2 6 0 0 n 4 for fetal bovine serum Invitrogen 1 4 0 1 for goat serum 1 3 0 1 for rat serum 2 9 0 1 for rat plasma 3 4 0 1 for human serum and 1 4 0 1 for human plasma respectively Uric Acid ODs90nm 0 10 20 30 Uric Acid mg dL Standard Curve in 96 well plate assay PUBLICATIONS 1 Viel E C et al 2008 Xanthine oxidase and mitochondria contribute to vascular superoxide anion generation in DOCA salt hypertensive rats Am J Physiol Heart Circ Physiol 295 H281 H288 2 Kamel A H 2007 Conventional and planar chip sensors for potentiometric assay of uric acid in biological fluids using flow injection analysis J Pharm Biomed Anal 45 2 341 348 3 DiSilvestro R A et al 2009 Pomegranate extract mouth rinsing effects on saliva measures relevant to gingivitis risk Phytother Res 23 8 1123 1127 Active Date 091 72012
4. ion is recommended Equilibrate to room temperature before assay Metal chelators e g EDTA interferew ith this assay and should be avoided Procedure using 96 well plate 1 Set up standards and samples Transfer 5 uL Blank Standard and samples in duplicate wells of a clear bottom 96 well plate 2 Add 200 uLworking reagent and tap lightly to mix 3 Incubate 30 min at room temperature and read optical density at 510 630nm peak absorbance at 590nm Procedure using cuvette 1 Set up test tubes labeled Blank Standard Samples Transfer 20 uL Blank Standard and samples to appropriately labeled tubes F 753 3UM RevA 25030021UA 2 Add 1000 pLworking reagent and tap lightly to mix 3 Incubate 30 min at room temperature and read optical density at 590nm 510nm 630nm CALCULATION The uric acid concentration of Sample is calculated as ODsampce ODsiank ODstanvarv ODstank X 10 mg dL ODerank ODstanparp and ODsampte are ODsgom values of Blank Standard and Sample respectively It is not necessary to prepare a calibration curve because the concentration of the provided standard lies w ithin the linear range Normal serum uric acid values 1 0 to 7 0 mg dL Conversions 1 mg dL uric acid equals 59 5 uM 0 001 or 10 ppm MATERIALS REQUIRED BUT NOT PROVIDED Pipeting devices and accessories e g 5 uL Procedure using 96 well plate Clear bottom 96 w ell plates e g Corning Costar 96 well plate absorbance
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