The RayBiotech Nitric Oxide (Nitrate/Nitrite) Assay Kit provides an accurate and convenient two-step process for measuring the total nitrate/nitrite concentration in serum, plasma, cell lysates, urine, or other biological fluid samples.
Nitric Oxide (Nitrate/Nitrite) Assay Kit
Nitric Oxide (Nitrate/Nitrite) Assay Kit
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Product Description
Specifications
| Size | 2 Plate Kit, 4 Plate Kit |
| Compatible Sample Types | Cell Culture Supernatants, Plasma, Serum, Tissue Lysates, Cell Lysates, Urine |
| Quantitative/Semi-Quantitative | Quantitative |
| Solid Support | 96-well Microplate |
| Method Of Detection | Colorimetric |
| Research Area | Cardiovascular Disease, Inflammation, Oxidative Stress |
| Shipping Type | Blue ice |
| Storage | 2-8°C |
| Storage / Stability | The entire kit should be stored at 2-8°C for up to 6 months from the date of shipment. Do not use the kit beyond its expiration date. For prepared reagent storage, see the manual. |
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Introduction
Nitric oxide (NO) is a highly reactive radical that plays an important role in many essential physiological functions, including neurotransmission, immune response, and apoptosis. It is produced through the oxidation of arginine via nitric oxide synthase, facilitating vital roles in host defense, activation of regulatory proteins, and direct covalent interactions with functional biomolecules. Given its inherent instability, NO undergoes rapid oxidative degradation to form nitrite (NO2-) and nitrate (NO3-). Therefore, quantifying the total concentration of NO2-/ NO3- is a reliable indicator of NO levels.
The RayBio Nitric Oxide (Nitrate/Nitrite) Assay Kit provides an accurate and convenient two-step process for measuring the total nitrate/nitrite concentration in serum, plasma, cell lysates, urine, or other biological fluid samples. First, nitrate is reduced to nitrite. Then, the addition of Griess reagent transforms nitrite into a deep purple azo compound. The nitrite concentration can be accurately determined by measuring the absorbance of the formed azo chromophore using spectrophotometry.
Kit Components
| Component | Size / Description |
|---|---|
| Microplates | Two 96-well plates (12 strips x 8 wells) |
| Sample Buffer | 10 ml |
| Acid Solution | 40 ml |
| Nitrate Standard | 1 vial (100μl) |
| Nitrite Standard | 1 vial (100μl) |
| Griess Reagent I | 1 bottle (powder) |
| Griess Reagent II | 1 bottle (powder) |
| Nitrate Reducing Reagent | 1 bottle (powder) |
Additional Materials Required
- Microplate reader capable of measuring absorbance at 540 nm
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Distilled or deionized water
- Tubes to prepare sample dilutions
- 50 ml conical tubes
- Incubator at 37°C.
- Centrifugal filter tubes with 10 or 30 kDa molecular weight cut-off
Assay Procedure Summary
- Prepare all reagents, nitrate/nitrite standards and samples as instructed.
- Add Nitrate/Nitrite working reagent.
- For Nitrate + Nitrite measurement, incubate the mixture for 30 minutes at 37°C. For Nitrite measurement, incubate the mixture for 10 minutes at room temperature.
- Transfer 200 μl of each reaction solution into separate wells of a 96-well plate.
- Read absorbance at 540 nm on a microplate reader.
documents
citations
"Oxidative Stress Indices and Inflammatory Responses in Female Wistar Rats Administered Acetaminophen, Ampicillin/Cloxacillin and Co-Trimoxazole"
- Species:
- Rat
- Year:
- 2021
- Journal Name:
- Biomedical Journal of Scientific & Technical Research
"Patterns of Some Immunological Proteins and Oxidative Indices in Normal and High Body Mass Index Wistar Rats Fed Coffee and Aspartame"
- Species:
- Rat
- Year:
- 2021
- Journal Name:
- Biomedical Journal of Scientific & Technical Research