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Xanthine Oxidase (XOD) Activity Assay Kit
SKU: E-BC-K805-M-96
Xanthine Oxidase (XOD) Activity Assay Kit
| SKU # | E-BC-K805-M |
| Detection Instrument | Microplate reader (540-560 nm, optimum wavelength: 550 nm) |
| Detection method: | Colorimetric method |
Product Details
Properties
| Synonyms | XOD |
| Sample type | Serum, plasma and animal tissue samples |
| Sensitivity | 0.067 U/L |
| Detection range | 0.067-39.30 U/L |
| Detection method |
Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 35 min |
| Precision | Average inter-assay CV: 9.900% | Average intra-assay CV: 3% |
| Other instruments required | Incubator(37℃) |
| Other reagents required | Normal saline (0.9% NaCl) |
| Storage | -20℃ |
| Valid period | 12 months |
Dilution of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Rat plasma | 1 |
| Rat serum | 1 |
| 10% Mouse kidney tissue homogenate | 1 |
| 10% Rat liver tissue homogenate | 1 |
| 10% Rat kidney tissue homogenate | 1 |
| 10% Mouse liver tissue homogenate | 1 |
Note: The diluent is normal saline (0.9% NaCl). For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
XOD can catalyze hypoxanthine to xanthine, and at the same time while producing superoxide anion free radicals. In the presence of electron acceptor and chromogenic agent, the purplish red substance can be generated, and the vitality of XOD can be calculated by measuring the production of purplish red substance.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 10 mL × 1 vial | 20 mL × 1 vial | -20℃, 12 months |
| Reagent 2 | Substrate Solution | 0.5 mL × 1 vial | 1 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 3 | Enzymatic Reagent | 0.5 mL × 1 vial | 1 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 4 | Chromogenic Agent 1 | 0.8 mL × 1 vial | 1.6 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 5 | Chromogenic Agent 2 | 0.8 mL × 1 vial | 1.6 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 6 | 1 mmol/L Standard Solution |
1.6 mL × 1 vial | 3.2 mL × 1 vial | -20℃, 12 months |
| 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 (U/L) | 1.20 | 13.50 | 26.40 |
| %CV | 3.4 | 3.0 | 2.6 |
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 (U/L) | 1.20 | 13.50 | 26.40 |
| %CV | 9.0 | 10.4 | 10.3 |
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 99%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mmol/L) | 0.25 | 0.46 | 0.77 |
| Observed Conc. (mmol/L) | 0.2 | 0.4 | 0.9 |
| Recovery rate(%) | 89 | 93 | 115 |
Sensitivity
The analytical sensitivity of the assay is 0.067 U/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 OD 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 (mmol/L) | 0 | 0.2 | 0.3 | 0.4 | 0.6 | 0.7 | 0.8 | 1 |
| OD value | 0.042 | 0.194 | 0.268 | 0.327 | 0.484 | 0.561 | 0.631 | 0.757 |
| 0.041 | 0.192 | 0.263 | 0.350 | 0.474 | 0.550 | 0.616 | 0.743 | |
| Average OD | 0.042 | 0.193 | 0.266 | 0.339 | 0.479 | 0.556 | 0.624 | 0.750 |
| Absoluted OD | 0 | 0.152 | 0.224 | 0.297 | 0.438 | 0.514 | 0.582 | 0.709 |