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Research Areas
Specifications
- Sample Water, Serum, Plasma, Urine, Saliva, Lysates, Buffers Tissue Culture Media
- Accurate Calibrated to NIST Standard Reference Material #3185
- Sensitivity 2.63 µM in the Nitrite and 1.02 µM in the Total Nitric Oxide - Highest Optical Density of Any Kit
- Time to Answer 5 Minute Nitrite – 25 Minute Total NO
- Samples/Kit 88 in duplicate
- Stability Non-Toxic, Liquid 4ºC stable reagents
- Readout Colorimetric, 550-570 nm
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Standard Curve
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Description
Nitric Oxide content is derived from the sum of Nitrate and Nitrite. Both Nitrate and Nitrite standards are provided to generate standard curves for the assay and all samples should be read off the appropriate standard curve. For Nitrite Detection, samples are mixed with the Color Reagents A and B and incubated at room temperature for 5 minutes. The colored product is read at 550–570 nm.
Total Nitric Oxide content is measured after the sample is incubated with Nitrate Reductase and NADH. The reductase in combination with NADH reduces Nitrate to Nitrite. After a 20 minute incubation at room temperature, Color Reagents A and B are added and incubated at room temperature for 5 minutes. The concentration of Nitrate in the sample is calculated by taking the measured Nitrite concentration and subtracting from the Total Nitric Oxide concentration for the sample.
Nitric oxide (NO) is a diffusible, transient, reactive molecule that has physiological effects in the pM-µM range. Acting through guanylate cyclase activation, NO is an important regulator of the cardiovascular, nervous, and immunological systems. NO is bio-available by two routes. It can be endogenously generated by constitutive or induced NOS enzymes, or it can be ingested as nitrates or nitrites for conversion into NO. The reactive nature of nitric oxide allows it to act as a cytotoxic factor when released during an immune response by macrophages. The reactivity also allows NO to be easily converted to a toxic radical that can produce nitrosylation damage to cells and DNA. Nitrosylation can be a regulated post-translational modification in cell signaling. The dynamics of the regulatory/damage facets of NO are major forces in mitochondrial signaling and dysfunction. NO is linked not only to coronary heart disease, endothelial dysfunctions, erectile dysfunction, and neurological disorders, but also diabetes, chronic periodontitis, autism and cancer.