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Nitric Oxide Assay Kit (Colorimetric) ab65328 provides an accurate, convenient measure of total nitrate/nitrite in a simple two-step process.

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

  • Nature communications 14:76392023
    Acoustofluidic Interfaces for the Mechanobiological Secretome of MSCs.
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Ye He,Shujie Yang,Pengzhan Liu,Ke Li,Ke Jin,Ryan Becker,Jinxin Zhang,Chuanchuan Lin,Jianping Xia,Zhehan Ma,Zhiteng Ma,Ruoyu Zhong,Luke P Lee,Tony Jun Huang
    PubMed 37993431
  • Nature communications 14:74272023
    Glutamine metabolism inhibition has dual immunomodulatory and antibacterial activities against Mycobacterium tuberculosis.
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Sadiya Parveen,Jessica Shen,Shichun Lun,Liang Zhao,Jesse Alt,Benjamin Koleske,Robert D Leone,Rana Rais,Jonathan D Powell,John R Murphy,Barbara S Slusher,William R Bishai
    PubMed 37973991

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 Clear Plate
2 x 1 Unit
Assay Buffer XVII
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 I
1 x 1 Vial
Nitrate Standard
1 x 1 Vial
Nitrite Standard
1 x 1 Vial

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Nitric Oxide Assay Kit (Colorimetric) ab65328 provides an accurate, convenient measure of total nitrate/nitrite in a simple two-step process.

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.

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 Griess reagent 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.

Supplementary info

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
    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)

    Nitrite and nitrate measured in biologicals showing concentration (micromolar)

  • 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