Nitric Oxide (NO) Colorimetric Detection Kit

Assay Principle:

The DetectX® Nitric Oxide (NO) Colorimetric Detection Kit quantitatively measures NO, nitrate, and nitrite levels in various samples, including water, serum, plasma, urine, saliva, lysates, buffers, and tissue culture media. This detection kit features a run time of 25 minutes, combining efficiency with precision. Read the complete kit insert for detailed instructions before starting the assay. The kit includes Nitrate and Nitrite standards to establish accurate standard curves.

Protocol Summary:

• Introduce standards or diluted samples into the provided transparent microtiter plate.
• Add Nitrate Reductase and NADH Concentrate to each well, ensuring thorough mixing.
• Incubate the mixture at room temperature for 20 minutes, allowing Nitrate to be reduced to Nitrite.
• Add Color Reagents A and B, initiating a color-generating reaction with the Nitrite in the sample.
• After a 5-minute incubation with the color reagents, use a plate reader to measure the signal at 540-570nm. Calculate NO concentration using the standard curve.

Background:

Nitric oxide (NO) is a versatile, transient, reactive molecule with significant physiological effects across a range of concentrations. It functions as a key regulator in the cardiovascular, nervous, and immunological systems, primarily through guanylate cyclase activation. NO is bio-available through endogenous generation by NOS enzymes and through ingestion as nitrates or nitrites, which are subsequently converted into NO.

NO’s reactive nature enables it to serve as a cytotoxic factor in immune responses. It is also a potentially damaging radical capable of inducing nitrosylation in cells and DNA. Nitrosylation can be both a harmful effect and a regulated post-translational modification in cell signaling. The interplay of NO’s regulatory and damaging roles is significant in mitochondrial signaling and dysfunction. NO influences a range of conditions, including coronary heart disease, endothelial dysfunctions, erectile dysfunction, neurological disorders, diabetes, chronic periodontitis, autism, and cancer. The DetectX® Nitric Oxide (NO) Colorimetric Detection Kit is an important tool for researchers exploring the diverse roles of NO in health and disease.