Description
Key facts
Detection method:Fluorescent,
Sample types:Urine, Plasma, Tissue Extracts, Serum, Other biological fluids, Cell Lysate,
Assay type:Quantitative,
Assay time:3h,
Assay Platform:Microplate reader
Product details:
Nitric Oxide Assay Kit (Fluorometric) ab65327 provides an accurate, convenient measure of total nitrate/nitrite concentration in a simple two-step process.
In the nitric oxide assay protocol:
- the first step converts nitrate to nitrite by nitrate reductase
- in the second step, nitrite reacts with the fluorescent probe DAN (2, 3 diaminonaphthalene)
The fluorescence can be measured at Ex/Em = 360/450 nm. NaOH enhances the fluorescent yield. The fluorescent intensity is proportional to the total nitric oxide production. The kit has been tested with culture media, plasma, and tissue homogenates.
Nitric oxide assay protocol summary:
- add samples and standards to wells
- add enzyme cofactor, to measure nitrite only add assay buffer or to measure total nitrate + nitrite add nitrate reductase
- incubate for 1-4 hr at room temp to convert nitrate to nitrite
- add enhancer and incubate for 30 min at room temp
- add DAN probe and incubate for 10 min at room temp
- add NaOH and incubate for 10 min at room temp
- analyze with microplate reader
This product is manufactured by BioVision, an Abcam company and was previously called K252 Nitric Oxide Fluorometric Assay Kit. K252-200 is the same size as the 200 test size of ab65327.
If you are interested in a colorimetric detection kit, please check Nitric Oxide Assay Kit (Colorimetric) (ab65328)
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. Since NO is rapidly converted to nitrite (NO2-) and nitrate (NO3-), the total concentration of nitrite and nitrate is used as a quantitative measure of NO production.
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-Please refer to protocols
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.
