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Lipoxygenase (LOX) Activity Assay Kit
SKU: E-BC-K861-M-96
Lipoxygenase (LOX) Activity Assay Kit
| SKU # | E-BC-K861-M |
| Detection Instrument | Microplate reader (270-290 nm, optimum wavelength: 280 nm) |
|
Detection method |
Colorimetric method |
Product Details
Properties
| Synonyms | LOX |
| Sample type | Plant samples |
| Sensitivity | 5 U/L |
| Detection range | 5-339.5 U/L |
| Detection method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 15 min |
| Precision | Average inter-assay CV: 5% | Average intra-assay CV: 4% |
| Other instruments required | Centrifuge, Incubator (37°C), Homogenizer |
| Other reagents required | Double distilled water, Tween-20 |
| Storage | 2-8℃ |
| Valid period | 12 months |
Dilution of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| 10% Long bean | 1 |
| 10% Green soy bean | 2-5 |
Note: The diluent is extraction solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
LOX can catalyze linoleic acid oxidation, the oxidation product has the specific absorption peak at 280 nm. The activity of LOX can be calculated by measuring the change of absorbance value at 280 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Extraction Solution | 50 mL × 1 vial | 50 mL × 2 vials | 2-8℃, 12 months |
| Reagent 2 | Buffer Solution | 15 mL × 1 vial | 30 mL × 1 vial | 2-8℃, 12 months |
| Reagent 3 | Substrate | 0.1mL × 1 vial | 0.1mL × 1 vial | 2-8℃, 12 months, shading light |
| UV 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 long bean 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) | 8.40 | 68.50 | 188.00 |
| %CV | 4.3 | 4.1 | 3.6 |
Inter-assay Precision
Three long bean 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) | 8.40 | 68.50 | 188.00 |
| %CV | 4.8 | 5.2 | 101 |
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%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/L) | 87.5 | 169 | 284 |
| Observed Conc. (U/L) | 86.6 | 163.9 | 286.8 |
| Recovery rate(%) | 99 | 97 | 101 |
Sensitivity
The analytical sensitivity of the assay is 0.036 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.