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Nitric Oxide (NO) Colorimetric Assay Kit (Nitrate Reductase Method)
SKU: E-BC-K135-M-96
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Nitric Oxide (NO) Colorimetric Assay Kit (Nitrate Reductase Method)
| SKU # | E-BC-K135-M |
| Detection Instrument | Microplate reader (520-550 nm, optimum wavelength: 530 nm) |
Product Details
Properties
| Synonyms | NO |
| Sample Type | Serum, plasma, urine, tissue, cell and supernatant of cell samples |
| Sensitivity | 1.38 μmol/L |
| Detection Range | 1.38-40 μmol/L |
| Detection Method | Quantitative |
| Assay time | 100 min |
| Precision | Average inter-assay CV: 8% | Average intra-assay CV: 6% |
| Other instruments required | Centrifuge, Vortex mixer, 37℃ Incubator |
| Storage | -20℃ |
| Valid period | 12 months |
Detection Principle
Nitric oxide (NO) has active chemical properties and is quickly metabolized into NO2- and NO3- in the body, while NO2- is further transformed into NO3-. This method uses nitrate reductase specifically to reduce NO3- to NO2-, and the concentration of NO is measured by the color depth.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Enzyme Reagent | Powder × 2 vials | Powder × 4 vials | -20℃, 12 months, shading light |
| Reagent 2 | Substrate | Powder × 1 vial | Powder × 1 vial | -20℃, 12 months, shading light |
| Reagent 3 | Sulfate Solution | 1.5 mL × 1 vial | 3 mL × 1 vial | -20℃, 12 months |
| Reagent 4 | Alkaline Reagent | 0.75 mL × 1 vial | 1.5 mL × 1 vial | -20℃, 12 months |
| Reagent 5 | Chromogenic Agent A |
Powder × 1 vial | Powder × 1 vial | -20℃, 12 months, shading light |
| Reagent 6 | Chromogenic Agent B |
Powder × 1 vial | Powder × 1 vial | -20℃, 12 months, shading light |
| Reagent 7 | Acid Reagent | 1.5 mL × 1 vial | 3 mL × 1 vial | -20℃, 12 months |
| Reagent 8 | 1 mmol/L Standard | 0.75 mL × 1 vial | 1.5 mL × 2 vials | -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 (μmol/L) | 5.50 | 18.50 | 32.00 |
| %CV | 6.5 | 5.8 | 5.7 |
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 (μmol/L) | 5.50 | 18.50 | 32.00 |
| %CV | 8.3 | 7.9 | 7.8 |
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. (μmol/L) | 12 | 22.5 | 31 |
| Observed Conc. (μmol/L) | 11.3 | 21.6 | 29.5 |
| Recovery rate (%) | 94 | 96 | 95 |
Sensitivity
The analytical sensitivity of the assay is 1.38 μmol/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 (μmol/L) | 0 | 8 | 16 | 20 | 24 | 28 | 32 | 40 |
| OD value | 0.043 | 0.092 | 0.144 | 0.166 | 0.194 | 0.214 | 0.249 | 0.295 |
| 0.043 | 0.089 | 0.143 | 0.165 | 0.194 | 0.214 | 0.250 | 0.300 | |
| Average OD | 0.043 | 0.091 | 0.144 | 0.166 | 0.194 | 0.214 | 0.250 | 0.298 |
| Absoluted OD | 0 | 0.048 | 0.101 | 0.123 | 0.151 | 0.171 | 0.207 | 0.255 |