Nitric Oxide Colorimetric Detection Kit
Referentie OKAU00021
Formaat : 2plate
Merk : Aviva Systems Biology
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| Datasheets/Manuals | Printable datasheet for Nitric Oxide Colorimetric Detection Kit (OKAU00021) |
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| Application | ELISA-Sandwich | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| ELISA Kit Detection Method | Colorimetric | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| ELISA Kit Linearity | Linearity was determined by taking two diluted human urine samples with known Nitrite and Total NO Concentrations and mixing them in the ratios given below. The measured concentrations were compared to the expected values based on the ratios used.
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| ELISA Kit Principle | The DetectX® Nitric Oxide Detection Kit is designed to quantitatively measure Nitrate and Nitrite present in a variety of samples. Nitric Oxide content is derived from the sum of Nitrate (- NO3 ) and Nitrite (- NO2 ). Please read the complete kit insert before performing this assay. 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 Color Reagents A and B and incubated at room temperature for 5 minutes. The colored product is read at 540 - 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. This kit uses Nitrate and Nitrite Standard solutions calibrated to the US National Institute for Science and Technology Standard Reference Materials and ISO/IEC standards. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| ELISA Kit Reproducibility | Intra Assay Precision Three samples were diluted in Assay Buffer and run in replicates of 20 in an assay. The mean and precision of the calculated Nitrite or Total NO concentrations were:
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| ELISA Kit Component |
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| Additional Information | Background: Nitric oxide (NO) is a diffusible, transient, reactive molecule that has physiological effects in the picomolarto-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 and 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. It 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 is also 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). | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| :: | Detection Limit: The Limit of Detection was determined as 0.94 uM in the Nitirite and 3.0 uM in the Total Nitric Oxide assays. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Reconstitution and Storage | 2°C to 8°C | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Sample Type | Water, Buffers, Serum, Plasma, Urine, Saliva, and TCM | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Sensitivity | Sensitivity was determined as 2.63 uM in the Nitrite and 1.02 uM in the Total Nitric Oxide assays. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Gene Full Name | Nitric Oxide Colorimetric |
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