Glycolate Oxidase Activity Assay Kit
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Glycolate Oxidase Activity Assay Kit
| SKU # | E-BC-K692-S |
| Detection Instrument | Spectrophotometer (324 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Sample type | Plant tissue |
| Sensitivity | 0.3 U/mL |
| Detection range | 0.3-350 U/mL |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 30 min |
| Precision | Average inter-assay CV: 8.200% | Average intra-assay CV: 1.100% |
| Other instruments required | Test tube, Micropipettor, Vortex mixer, Optical path cuvette (1 mL of volume, 1 cm optical diameter) |
| Storage | 2-8℃ |
| 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 (0.3-350 U/mL).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| 10% Oxalis corniculata tissue homogenate | 1 |
| 10% Ginkgo biloba tissue homogenate | 1 |
| 10% Cactus tissue homogenate | 1 |
| 10% Bamboo leaf tissue homogenate | 1 |
| 10% Osmanthus fragrans leaf tissue homogenate | 4-8 |
| 10% Camphor trees leaf tissue homogenate | 3-5 |
Note: The diluent is reagent 1.
Detection Principle
Glycolate oxidase catalyzes sodium glycolate substrate to form glyoxylic acid, which reacts with phenylhydrazine hydrochloride to form phenylhydrazone glyoxalate. The substance has an absorption peak at 324 nm, and its OD value is proportional to the concentration of phenylhydrazone glyoxalate in a certain range, and the amount of phenylhydrazone generated reflects the activity of glycolate oxidase.
Kit Components & Storage
| Item | Component |
Size 1 (50 assays) |
Size 2 (100 assays) |
Storage |
| Reagent 1 | Extraction Solution | 60 mL × 1 vial | 60 mL × 2 vials | 2-8℃, 12 months |
| Reagent 2 | Buffer Solution | 45 mL × 1 vial | 45 mL × 2 vial | 2-8℃, 12 months shading light |
| Reagent 3 | Chromogenic Agent | Powder × 1 vial | Powder × 1 vial | 2-8℃, 12 months, shading light |
| Reagent 4 | Substrate | 6 mL × 1 vial | 12 mL × 1 vial | 2-8℃, 12 months |
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 cactus tissue 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/mL) | 5.80 | 85.50 | 179.00 |
| %CV | 1.3 | 1.2 | 0.8 |
Inter-assay Precision
Three cactus tissue samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (U/mL) | 5.80 | 85.50 | 179.00 |
| %CV | 8.0 | 8.1 | 8.5 |
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 102%.
Sensitivity
The analytical sensitivity of the assay is 0.3 U/mL. 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.