Nitric Oxide Detection Kit

$360.00

The Nitric Oxide Detection Kit is designed to quantitatively measure Nitrate and Nitrite present in a variety of samples.

SKU: SKT-212-96 Categories: , ,

Nitric Oxide Detection Kit

The Nitric Oxide Detection Kit is For Research Use Only

Size: 1×96 wells
Sensitivity: 2.63 µM in the Nitrite and 1.02 µM in the Total Nitric Oxide
Dynamic Range: 3.125 – 200 µM
Incubation Time: 30 minutes
Sample Type: Buffer, Cell lysates, Plasma, Saliva, Serum, Tissue Culture Media, Urine, Water
Sample Size: 50 μL

Product manufactured in Canada by StressMarq.


Assay Background

Nitric oxide (NO) is a diffusible, transient, reactive molecule that has physiological effects in the picomolar-to-micromolar range. Acting through soluble guanylate cyclase activation, NO is an important physiological regulator of the cardiovascular, nervous, and immunological systems. NO is bio-available by two routes. It can be endogenously generated by constitutive or induced enzymes like Nitric Oxide Synthase or it can be orally ingested as nitrates / nitrites for rapid uptake into circulation and subsequent conversion. The reactive nature of nitric oxide allows it to act as a cytotoxic factor when released during an immune response by cells such as macrophages. The reactivity also allows NO to be easily converted to a toxic radical that can produce nitrosative damage to cells, organelles and molecules such as DNA. Nitrosaylation however can be a regulated post-translational modification in cell signaling. The balance and 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, cancer, and assorted age-related diseases. The physical properties of Nitric Oxide make it challenging for direct detection methods. However, colorimetric methods can be applied to measure its stable break-down products nitrate (-NO3) and nitrite (-NO2).


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Additional Information

Assay Principle


Nitric Oxide content is derived from the sum of Nitrate (-NO3) and Nitrite (-NO2). 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. The concentration of Nitrite in the sample is calculated, after making a suitable correction for any dilution of the sample, using software available with most plate readers. 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 colored product is read and calculated as with the Nitrite determination above. The concentration of Nitrate in the sample is calculated by subtracting the measured Nitrite concentration from the Total Nitric Oxide concentration for the sample.

Typical Standard Curve


Nitric Oxide Detection Kit

Manual

Product Manual