Abcam, ab65328, Nitric Oxide Assay Kit (Colorimetric)

Size: 2 x 96Tests
Nitric Oxide Assay Kit (Colorimetric) ab65328 uses nitrate reductase and the Griess reagent method. It can be used to quantify both total nitrate/nitrite levels and/or nitrate and nitrite levels separately. Readout on any colorimetric (540 nm) plate reader. - Cited in >120 publications - Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Key facts
Detection method:Colorimetric,
Sample types:Urine, Plasma, Tissue Extracts, Serum, Other biological fluids, Cell Lysate,
Assay type:Quantitative,
Sensitivity:= 1 nmol/well,
Range:1 - 10 nmol/well,
Assay time:1h 30m,
Assay Platform:Microplate reader

Product details:
Nitric Oxide Assay Kit (Colorimetric) ab65328 provides an accurate, convenient measure of total nitrate/nitrite in a simple two-step process.
Nitric oxide assay principle
- 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 (540 nm).
The amount of the azochromophore accurately reflects the nitric oxide amount in samples.
The assay measures total nitrate + nitrite levels. To measure nitrate and nitrite levels separately, samples can also be run without the nitrate reductase, to measure nitrite, and nitrate levels are calculated by subtracting nitrite levels from total nitrate + nitrite.
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
Related and recommended products
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.
This product was previously called K262 Biovision Nitric Oxide Colorimetric Assay Kit. Biovision was acquired by Abcam in 2021.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Nitric oxide (NO) is a small diatomic molecule with a mass of 30.01 Da. It plays a significant mechanical role as a signaling molecule in numerous systems. Nitric oxide is synthesized by nitric oxide synthases (NOS) and is expressed in endothelial cells neurons and various immune cells. Its production occurs in response to mechanical shear stress calcium influx or immunological stimulation. Nitric oxide exhibits high diffusibility enabling it to cross cellular membranes easily which is key for its function as a signaling messenger.
Biological function summary
Nitric oxide mediates vasodilation neurotransmission and immune response modulation. It is important for maintaining vascular homeostasis where it relaxes smooth muscle tissue promoting blood flow. In the central nervous system nitric oxide acts as a neurotransmitter and neuromodulator influencing synaptic plasticity. Within the immune system it functions to regulate leukocyte adhesion and can mediate microbial destruction. Nitric oxide often works within complexes associating with proteins like guanylate cyclase enhancing its signaling efficacy through nitrosylation.
Pathways
Nitric oxide integrates into key biological processes such as the nitric oxide signaling pathway and the endothelial nitric oxide pathway. It activates soluble guanylate cyclase increasing cyclic GMP levels leading to smooth muscle relaxation and vasodilation. In the endothelial pathway nitric oxide links with other proteins like endothelium-derived relaxing factor and interacts with oxidative stress pathways showing its role in cellular defense mechanisms. These pathways manage critical physiological responses and connect nitric oxide to other cardiovascular regulators.
Nitric oxide dysregulation associates with conditions like cardiovascular disease and neurodegenerative disorders. A deficiency or excess production can contribute to hypertension or atherosclerosis given its role in vascular health. Neurodegenerative diseases such as Alzheimer's disease also show links to nitric oxide largely due to its involvement in neuroinflammation and synaptic dysfunctions. It interacts with proteins like amyloid-beta precursor protein and tau in these contexts highlighting its importance in pathology and therapeutic targets.