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Oxalate (Oxalic Acid) Colorimetric Assay Kit
SKU: E-BC-K892-M-500
Oxalate (Oxalic Acid) Colorimetric Assay Kit
| SKU # | E-BC-K892-M |
| Detection Instrument | Micropipettor, Microplate reader (545-555 nm, optimum wavelength: 550 nm) |
|
Detection method |
Colorimetric method |
Product Details
Properties
| Sample type | Animal urine, serum, plasma, plant tissue |
| Sensitivity | 0.02 mmol/L |
| Detection range | 0.02-1 mmol/L |
| Detection method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 35 min |
| Precision | Average inter-assay CV: 4.700% | Average intra-assay CV: 3% |
| Other instruments required | Micropipettor, Centrifuge, 37℃ incubator |
| Storage | -20℃ |
| Valid period | 12 months |
Dilution of Sample
It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range ( 0.02-1 mmol/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human urine | 1-2 |
| Human plasma | 1 |
| 10% Epipremnum aureum tissue homogenate | 2-3 |
| Rats plasma | 1 |
Note: The diluent is double distilled water.
Detection Principle
Oxalate oxidase catalyzes the oxidation of oxalate to produce hydrogen peroxide and carbon dioxide. Under the action of POD, hydrogen peroxide reacts with chromogenic substances to produce colored products. There is a specific absorption peak at 550 nm, and the color depth is proportional to the content of oxalate.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Working Solution | 12 mL × 1 vial | -20℃, 12 months |
| Reagent 2 | Chromogenic Agent A | 12 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 3 | Chromogenic Agent B | 12 mL × 1 vial | -20℃, 12 months, shading light |
| Reagent 4 | Enzyme Reagent A | Powder × 2 vials | -20℃, 12 months |
| Reagent 5 | Enzyme Reagent B | Powder × 2 vials | -20℃, 12 months |
| Reagent 6 | Regulator | 12 mL × 1 vial | -20℃, 12 months |
| Reagent 7 | Clarificant | 2 mL × 1 vial | -20℃, 12 months |
| Reagent 8 | 1 mmol/L Standard | 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 (mmol/L) | 0.08 | 0.54 | 0.88 |
| %CV | 3.5 | 3.1 | 2.4 |
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 (mmol/L) | 0.08 | 0.54 | 0.88 |
| %CV | 4.8 | 5.2 | 4.1 |
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 95%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (mmol/L) | 0.35 | 0.55 | 0.75 |
| Observed Conc. (mmol/L) | 0.3 | 0.5 | 0.7 |
| Recovery rate(%) | 96 | 94 | 95 |
Sensitivity
The analytical sensitivity of the assay is 0.02 mmol/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 | 0.4 | 0.5 | 0.6 | 0.7 | 0.8 | 1.0 |
| Average OD | 0.046 | 0.137 | 0.228 | 0.268 | 0.316 | 0.353 | 0.390 | 0.468 |
| Absoluted OD | 0.000 | 0.090 | 0.182 | 0.222 | 0.270 | 0.307 | 0.344 | 0.422 |