Nitric Oxide Detection Kit
Based on 1 Customer Validation
MCE Nitric Oxide Detection Kit employs a modified Griess method to detect the content of Nitric Oxide in various samples.
-
Storage :
-20℃, 1 year
Keep away from light and avoid repeated freezing and thawing
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Description & Advantages
Nitric oxide (NO) is a reactive radical, an oxidation product of arginine by nitric oxide synthase. NO is involved in host defense and development, activation of regulatory proteins and direct covalent interaction with functional biomolecules, plays an important role in many key physiological functions.
MCE Nitric Oxide Detection Kit employs a modified Griess method to detect the content of Nitric Oxide in various samples. The detection principle is that Nitric Oxide is easily oxidized to form Nitrite (NO2-), and by measuring the Nitrite content, the Nitric Oxide content in the sample can be determined.
Features of MCE Nitric Oxide Detection Ki
1. Excellent Linearity: Displays good linearity within the range of 1-100 μM.
2. Simple and fast: It only takes 5-10 min to complete the standard curve drawing and sample testing.
3. Broad Sample Applicability: Suitable for detecting Nitric Oxide content in a variety of samples, including plasma, serum, urine, tissues, and cells. The presence of phenol red and 10% serum does not significantly influence the results.
Protocol
Standard Curve Preparation
1. Dilute NaNO2 (1 M) with deionized water to prepare a 1 mM intermediate solution.
2. Using the same culture medium, PBS, or other suitable diluent as used for samples, prepare 0, 1, 2, 5, 10, 20, 50, and 100 μM NaNO2 standards.
Sample Preparation
1. Cell-culture supernatant, plasma, serum, or urine may be assayed directly or diluted as required. Centrifuge turbid samples and use the clear supernatant.
2. Prepare cell or tissue homogenates/lysates with nitrite-free buffer, centrifuge at 10,000 × g for 10 min at 4°C, and collect the supernatant. Deproteinize if necessary.
Assay Procedure
| Well | Sample/Standard | Griess Reagent I | Griess Reagent II |
|---|---|---|---|
| Blank | 50 μL diluent | 50 μL | 50 μL |
| Standard | 50 μL NaNO2 standard | 50 μL | 50 μL |
| Sample | 50 μL sample | 50 μL | 50 μL |
1. Add blanks, standards, and samples to a clear 96-well plate according to the table.
2. Add 50 μL Griess Reagent I to each well, mix gently, and incubate at room temperature for 5 min.
3. Add 50 μL Griess Reagent II to each well, mix gently, and incubate at room temperature for 5-10 min protected from light.
4. Measure absorbance at 540 nm. If a 540 nm filter is unavailable, use the closest wavelength between 520 and 560 nm.
Calculation
Subtract the blank absorbance, generate a NaNO2 standard curve, and determine nitrite concentrations in samples as an index of NO level. Multiply by the sample dilution factor.
Storage
-20℃, 1 year
Keep away from light and avoid repeated freezing and thawing
Components
| Components | HY-K0318-100 T |
|---|---|
| NaNO2 (1 M) | 200 μL |
| Griess Reagent I | 5 mL |
| Griess Reagent II | 5 mL |
Please enter the correct Lot Number
