Skip to main content

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.


Images

Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (AB65328), expandable thumbnail
  • Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (AB65328), expandable thumbnail
  • Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (AB65328), expandable thumbnail
  • Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (AB65328), expandable thumbnail
  • Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (AB65328), expandable thumbnail

Publications

Key facts

Detection method
Colorimetric
Sample types
Urine, Plasma, Tissue Extracts, Serum, Other biological fluids, Cell Lysate
Assay type
Quantitative
Reactive species
Mammals
Range
1 - 10 nmol/well
Assay time
1h 30m
Sensitivity
= 1 nmol/well

Associated Products

Select an associated product type

2 products for Alternative Product

What's included?

2 x 96 Tests
Components
96-Well Strip Clear Plates
2 x 1 Unit
Assay Buffer 17
1 x 30 mL
Enhancer II
1 x 1 Vial
Enzyme Cofactor
1 x 1 Vial
Griess Reagent I
1 x 10 mL
Griess Reagent II
1 x 10 mL
Microplate Sealing Film
2 x 1 Unit
Nitrate Reductase Enzyme
1 x 1 Vial
Nitrate Standard
1 x 1 Vial
Nitrite Standard
1 x 1 Vial

Recommended products

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
Reactive species
Mammals
Range
1 - 10 nmol/well
Assay time
1h 30m
Assay Platform
Microplate reader
Sensitivity
= 1 nmol/well

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C

Notes

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) Nitric Oxide Assay Kit (Fluorometric) ab65327.
For an assay kit to detect nitrite alone see Griess Reagent Kit Griess Assay Kit / 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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

5 product images

  • Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (ab65328), expandable thumbnail

    Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (ab65328)

    Nitrite and nitrate measured in biologicals showing concentration (micromolar)

  • Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (ab65328), expandable thumbnail
    Nam, Eunryel et al., Scientific reports vol. 8,1 7312., Fig 4 , doi:10.1038/s41598-018-25786-x

    Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (ab65328)

    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.

  • Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (ab65328), expandable thumbnail

    Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (ab65328)

    Nitrite and nitrate measured in cell lysates showing quantity (micromol) per 106 cells

  • Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (ab65328), expandable thumbnail

    Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (ab65328)

    Standard curve (colorimetric) : mean of duplicates (+/-SD) with background readings subtracted

  • Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (ab65328), expandable thumbnail

    Functional Studies - Nitric Oxide Assay Kit (Colorimetric) (ab65328)

    Standard curve (Colorimetric) : mean of duplicates (+/-SD) with background readings subtracted

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com