Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 1 hr 30 min
- Sample type: Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
- Sensitivity: 1 nmol/well
Product nameNitric Oxide Assay Kit (Colorimetric)
See all Nitric oxide kits
Sample typeUrine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate
Range1 nmol/well - 10 nmol/well
Assay time1h 30m
Species reactivityReacts with: Mammals, Other species
Nitric Oxide Assay Kit (Colorimetric) ab65328 provides an accurate, convenient measure of total nitrate/nitrite in a simple two-step process.
In the nitric oxide assay protocol:
- the first step converts nitrate to nitrite by using nitrate reductase
- the second step uses Griess Reagent to convert nitrite to a deep purple azo compound.
The amount of the azochromophore accurately reflects nitric oxide amount in samples.
Nitric oxide assay protocol summary:
- add samples and standards to wells
- add nitrate reductase and enzyme cofactor and incubate for 1 hr at room temp to convert nitrate to nitrite
- add enhancer and incubate for 10 min at room temp
- add Griess Reagent R1 and Griess Reagent R2
- analyze with microplate reader
This product is manufactured by BioVision, an Abcam company and was previously called K262 Nitric Oxide Colorimetric Assay Kit. K262-200 is the same size as the 2 96 tests size of ab65328.
If you are interested in a fluorometric detection kit, please check Nitric Oxide Assay Kit (Fluorometric) (ab65327).
For an assay kit to detect nitrite alone see Griess Reagent Kit ab234044.
Nitric oxide (NO) plays an important role in neurotransmission, vascular regulation, immune response and apoptosis. NO is rapidly oxidized to nitrite and nitrate which are used to quantitate NO production.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 2 x 96 tests Assay Buffer XVII 1 x 30ml Enhancer II 1 vial Enzyme Cofactor 1 vial Griess Reagent I 1 x 10ml Griess Reagent II 1 x 10ml 96-Well Clear Plate 2 units Nitrate Reductase I 1 vial Nitrate Standard 1 vial Nitrite Standard 1 vial TEMPPLATE® SEALING FILM 2 units
RelevanceNitric oxide (NO) is a key vertebrate biological messenger, playing an important role in neurotransmission, vascular regulation, immune responses and apoptosis. NO , also known as "endothelium-derived relaxing factor" or "EDRF", is synthesized from L-arginine, oxygen and NADPH by various NO synthases. Most of the NO in the cell is oxidized to nitrite (NO2-) and nitrate (NO3-), and therefore the concentrations of these anions are generally as a quantitative measure of NO production.
Blood NOx concentration measured with ab65328. Blood NOx concentration after application of the NE with NO. The level of blood NOx concentration increased 3-fold after the application of the NE with NO compared with control group.
Nitrite and nitrate measured in cell lysates showing quantity (micromol) per 106 cells
Nitrite and nitrate measured in biologicals showing concentration (micromolar)
Standard curve (colorimetric) : mean of duplicates (+/-SD) with background readings subtracted
Standard curve (Colorimetric) : mean of duplicates (+/-SD) with background readings subtracted
ab65328 has been referenced in 99 publications.
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- Lau M et al. Enhanced Antioxidant Effects of the Anti-Inflammatory Compound Probucol When Released from Mesoporous Silica Particles. Pharmaceutics 14:N/A (2022). PubMed: 35335878
- Mangmool S et al. Anti-Inflammatory Effects of Mitrephora sirikitiae Leaf Extract and Isolated Lignans in RAW 264.7 Cells. Molecules 27:N/A (2022). PubMed: 35630789