NADH Oxidase (NOX) Activity Assay Kit
NADH Oxidase (NOX) Activity Assay Kit
| SKU # | E-BC-K806-M |
| Detection Instrument | Microplate reader (590-610 nm, optimum wavelength: 600 nm) |
| Detection method: | Colorimetric method |
Product Details
Properties
| Synonyms | NOX |
| Sample type | Plant, animal tissue and cell samples |
| Sensitivity | 0.38 U/L |
| Detection range | 0.38-22.09 U/L |
| Detection method |
Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 15 min |
| Precision | Average inter-assay CV: 7.100% | Average intra-assay CV: 4% |
| Other instruments required | Centrifuge |
| 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 |
| 10% Mouse liver tissue homogenate | 5-10 |
| 10% Mouse heart tissue homogenate | 2-3 |
| 10% Porcine heart tissue homogenate | 1-3 |
| 10% Rat brain tissue homogenate | 1 |
| 10% Mouse kidney tissue homogenate | 3-5 |
| 10% Mouse muscle tissue homogenate | 1 |
| 10% Bovine liver tissue homogenate | 5-8 |
| 10%Epipremnum aureum tissue homogenate | 1 |
Note: The diluent is extraction solution B. For the dilution of other sample types, please do pretest to confirm the dilution factor
Detection Principle
NADH Oxidase（NOX）is widely found in the animals, plants, microorganisms and cultured cells, which can directly oxidize NADH to NAD+ in the presence of oxygen, and reduce blue DCPIP to colorless DCPIP. The activity of NOX can be calculated by measuring the reduction rate of blue DCPIP at 600 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Extraction Solution A | 50 mL × 1 vial | 50 mL × 2 vials | -20℃, 12 months |
| Reagent 2 | Extraction Solution B | 15 mL × 1 vial | 30 mL × 1 vial | -20℃, 12 months |
| Reagent 3 | Inhibitor | 0.8 mL × 1 vial | 0.8 mL × 2 vials | -20℃, 12 months, shading light |
| Reagent 4 | Buffer Solution | 10 mL×1 vial | 20 mL ×1 vial | -20℃, 12 months |
| Reagent 5 | Substrate A | 1.2 mL×1 vial | 1.2 mL ×2 vials | -20℃, 12 months, shading light |
| Reagent 6 | Substrate B | Powder × 1 vial | Powder ×2 vials | -20℃, 12 months, shading light |
| 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 mouse heart 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/L) | 2.60 | 13.50 | 18.40 |
| %CV | 4.3 | 4.0 | 3.7 |
Inter-assay Precision
Three mouse heart 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/L) | 2.60 | 13.50 | 18.40 |
| %CV | 6.8 | 7.0 | 7.5 |
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%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/L) | 5.2 | 10.8 | 16.5 |
| Observed Conc. (U/L) | 5.1 | 11.1 | 17.2 |
| Recovery rate(%) | 99 | 103 | 104 |
Sensitivity
The analytical sensitivity of the assay is 0.38 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.