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Nitrite Colorimetric Assay Kit
SKU: E-BC-K070-S-100
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Nitrite Colorimetric Assay Kit
| SKU # | E-BC-K070-S |
| Detection Instrument | Spectrophotometer (550 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Sample Type | Serum, plasma, saliva, tissue, cells, cell culture supernatant |
| Sensitivity | 1.36 μmol/L |
| Detection Range | 1.36-500 μmol/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 60 min |
| Precision | Average inter-assay CV: 3.300% | Average intra-assay CV: 2.600% |
| Other instruments required | Micropipettor, Vortex mixer, Centrifuge |
| Other reagents required | Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| Storage | Reagent 6: -20℃, others: 2-8℃ |
| Valid period | 12 months |
Detection Principle
Nitrite can react with chromogenic agent producing light red azo-compound. The content of nitrite can be calculated indirectly by measuring the OD value at 550 nm.
Kit Components & Storage
| Item | Component | Size 1 (50 assays) | Size 2 (100 assays) | Storage |
| Reagent 1 | Saline Solution | 50 mL ×1 vial | 50 mL ×2 vials | 2-8℃, 12 months |
| Reagent 2 | Alkali Reagent | 25 mL ×1 vial | 50 mL × 1 vial | 2-8℃, 12 months |
| Reagent 3 | Chromogenic Agent A | Powder × 1 vial | Powder ×1 vial | 2-8℃, 12 months shading light |
| Reagent 4 | Chromogenic Agent B | Powder × 1 vial | Powder ×1 vial | 2-8℃, 12 months shading light |
| Reagent 5 | Acid Solution | 6 mL ×1 vial | 12 mL × 1 vial | 2-8℃, 12 months |
| Reagent 6 | Sodium Nitrite Standard | Powder × 1 vial | Powder × 1 vial | -20℃, 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 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) | 12.80 | 156.00 | 368.00 |
| %CV | 2.8 | 2.4 | 2.6 |
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) | 12.80 | 156.00 | 368 |
| %CV | 3.1 | 3.2 | 3.6 |
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 97%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (μmol/L) | 88.5 | 264 | 403 |
| Observed Conc. (μmol/L) | 87.6 | 256.1 | 382.9 |
| Recovery rate (%) | 99 | 97 | 95 |
Sensitivity
The analytical sensitivity of the assay is 1.36 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 | 20 | 60 | 100 | 200 | 300 | 400 | 500 |
| Average OD | 0.002 | 0.002 | 0.002 | 0.002 | 0.002 | 0.002 | 0.002 | 0.002 |
| Absoluted OD | 0.051 | 0.051 | 0.051 | 0.051 | 0.051 | 0.051 | 0.051 | 0.051 |