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Uric Acid (UA) Fluorometric Assay Kit
SKU: E-BC-F018-96
Uric Acid (UA) Fluorometric Assay Kit
| SKU # | E-BC-F018 |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=535 nm/587 nm) |
| Detection Method | Fluorescence method |
Product Details
Properties
| Synonyms | UA |
| Sample Type | Serum, plasma, urine, animal tissue |
| Sensitivity | 0.03 μmol/L |
| Detection Range | 0.03-15 μmol/L |
| Detection Method | Fluorescence method |
| Assay Time | 40 min |
| Precision | Inter-assay CV: 7.2% | intra-assay CV: 1.5% |
| Storage | This product can be stored at -20°C for 12 months with shading light. |
| Valid period | 12 months |
Dilution Of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
|---|---|
| Human serum | 10-20 |
| Human urine | 80-100 |
| Human hydrothorax | 50-60 |
| Rat urine | 10-20 |
| Rabbit serum | 5-10 |
| Rat serum | 10-20 |
| Porcine serum | 1 |
| 10% Rat liver tissue homogenate | 10-20 |
| 10% Rat kidney tissue homogenate | 30-40 |
| 10% Rat lung tissue homogenate | 10-20 |
The diluent is buffer solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Uricase catalyzes the decomposition of uric acid into allantoin, CO2 and H2O2. Under the action of peroxidase, H2O2 oxidizes the non-fluorescent probe into the fluorescent substance. By measuring the fluorescence value of the system, the corresponding uric acid content can be calculated.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 60 mL × 1 vial | 60 mL × 2 vials | -20℃, 12 months |
| Reagent 2 | Probe Solution | 0.12 mL × 1 vial | 0.24 mL × 1 vial -2 | 2-8°C, 12 months shading light |
| Reagent 3 | Enzyme Reagent 1 | 0.12 mL × 1 vial | 0.24 mL × 1 vial | -20℃, 12 months |
| Reagent 4 | Enzyme Reagent 2 | 0.6 mL × 1 vial | 1.2 mL × 1 vial | -20℃, 12 months |
| Reagent 5 | 20 μmol/L Uric Acid Standard | 1.5 mL × 1 vial | 1.5 mL × 1 vial | -20℃, 12 months |
| Black Microplate | 96 wells | No requirement | ||
| Plate Sealer | 2 pieces | |||
Note: The reagents must be stored strictly according to the preservation conditions
in the above table. The reagents in different kits cannot be mixed with each other.
For a small volume of reagents, please centrifuge before use, so as not to obtain
sufficient amount of reagents.
Technical Data:
Parameter:
Intra-assay Precision
Three human serum samples were assayed in replicates of 20 to determine precision within an assay. (CV = Coefficient of Variation)
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol/L) | 0.20 | 3.50 | 9.80 |
| %CV | 2.0 | 1.2 | 1.3 |
Inter-assay Precision
Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol/L) | 0.20 | 3.50 | 9.80 |
| %CV | 7.9 | 6.8 | 6.9 |
Recovery
Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 101%
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (μmol/L) | 2.3 | 7.5 | 11.6 |
| Observed Conc. (μmol/L) | 2.3 | 7.4 | 12.1 |
| Recovery rate (%) | 100 | 99 | 104 |
Sensitivity
The analytical sensitivity of the assay is 0.03 μmol/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.
Standard Curve:
As the fluorescence value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only：
| Concentration (μmol/L) | 0 | 2 | 4 | 6 | 8 | 10 | 12 | 15 |
| Fluorescence value | 269 | 908 | 1758 | 2466 | 2974 | 3633 | 4263 | 4956 |
| 263 | 949 | 1832 | 2626 | 3212 | 3921 | 4701 | 5285 | |
| Average fluorescence value | 266 | 929 | 1805 | 2546 | 3093 | 3777 | 4482 | 5121 |
| Absoluted fluorescence value | 0 | 663 | 1539 | 2280 | 2827 | 3511 | 4216 | 4855 |