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Total NO/NO2-/NO3- S (1 PACK)
品牌： R&D Systems
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Product Summary
The Parameter Total Nitric Oxide kit has two assay options. Endogenous nitrite is measured in the first option. In the second option, nitrate is converted to nitrite using nitrate reductase and total nitrite is measured. To obtain the nitrate concentration, endogenous nitrite is subtracted from the total nitrite value.
Recovery
The recovery of the nitrate standard spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Supernates (Nitrite Assay) (n=10) | 107 | 91-115 |
| Plasma (Nitrate Assay) (n=25) | 98 | 87-110 |
| Plasma (Nitrite Assay) (n=20) | 104 | 92-117 |
| Serum (Nitrate Assay) (n=10) | 95 | 89-101 |
| Serum (Nitrite Assay) (n=10) | 109 | 98-118 |
| Urine (Nitrite Assay) (n=10) | 101 | 87-112 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of the nitrate standard were serially diluted with the Reaction Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data
Assay Procedure
Nitrite Assay ProcedureNitrate Reduction Assay Procedure
Refer to the product
This assay procedure measures the concentration of endogenous nitrite present in the sample. Bring all reagents and samples to room temperature before use. It is recommended that all samples and standards be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Reaction Diluent (1X) to the Blank wells.
- Add 50 µL of Nitrite Standard or sample to the remaining wells.
- Add 50 µL of Reaction Diluent (1X) to all wells.
- Add 50 µL of Griess Reagent I to all wells.
- Add 50 µL of Griess Reagent II to all wells. Mix by gently tapping the side of the plate.
- Incubate at room temperature for 10 minutes. Read at 540 nm with wavelength correction at 690 nm.
50 µL Reaction Diluent (1X)
50 µL Nitrite Standard or Sample
50 µL Reaction Diluent (1X)
50 µL Griess Reagent I
50 µL Griess Reagent II
Refer to the product
This assay procedure measures total nitrite by converting nitrate to nitrite. To determine the nitrate concentration in the sample, the endogenous nitrite concentration measured from the Nitrite Assay Procedure must be subtracted from the converted nitrite concentration measured in this procedure. Bring all reagents and samples to room temperature before use. It is recommended that all samples and standards be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Reaction Diluent (1X) to the Blank wells.
- Add 50 µL of Nitrate Standard or sample to the remaining wells.
- Add 25 µL of NADH to all wells.
- Add 25 µL of diluted Nitrate Reductase to all wells. Mix well, cover with a plate sealer, and incubate at 37 °C for 30 minutes.
- Add 50 µL of Griess Reagent I to all wells.
- Add 50 µL of Griess Reagent II to all wells. Mix by gently tapping the side of the plate.
- Incubate at room temperature for 10 minutes. Read at 540 nm with wavelength correction at 690 nm.
50 µL Reaction Diluent (1X)
50 µL Nitrate Standard or Sample
25 µL NADH
25 µL Nitrate Reductase
50 µL Griess Reagent I
50 µL Griess Reagent II
Total Nitric Oxide and Nitrate/Nitrite Parameter Assay Kit Summary
Format
96-well strip plate
Assay Length
< 1 hour
Sample Type & Volume Required Per Well
Cell Culture Supernates (10 uL), Serum (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL), Citrate Plasma (25 uL), Urine (10 uL)
Sensitivity
0.78 umol/L
Assay Range
3.1 - 200 umol/L (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
Specificity
Measures Total Nitric Oxide, Nitrite and Nitrate levels in various samples
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
Interference observed with other substances.
背景
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保存方式
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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