StressXpress®
Nitric Oxide Detection Kit (Discontinued)

Colorimetric detection of nitrate and nitrite

Catalog No. SKT-212

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Species Reactivity ALL
Sample Types Buffer, Cell lysates, Plasma, Saliva, Serum, Tissue Culture Media, Urine, Water
SKU: SKT-212 Categories: ,

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SKT-212_Nitric_Oxide_Detection_Kit_Standard_Curve_Fig1.png
Graph of the Linearity Recovery for the Nitric Oxide Detection Kit StressXpress - SKT-212
Product Name Nitric Oxide Detection Kit (Discontinued)
Description

Colorimetric detection of nitrate and nitrite

Species Reactivity Species Independent
Platform Microplate
Sample Types Buffer, Cell lysates, Plasma, Saliva, Serum, Tissue Culture Media, Urine, Water
Detection Method Colorimetric Assay
Assay Type Indirect Quantitative Assay
Utility Colorimetric assay used to quantitatively measure Nitrate and Nitrite present in a variety of samples.
Sensitivity 2.63 µM in the Nitrite and 1.02 µM in the Total Nitric Oxide
Assay Range 3.125 - 200 µM
Precision Intra Assay Precision: Three samples were further 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: Sample 1 (Nitrite Concentration)- 45.1 µM, 4.4% CV, Sample 1 (Total NO Concentration)- 70.5 µM, 6.8% CV Sample 2 (Nitrite Concentration)- 73.3 µM, 9.1% CV, Sample 1 (Total NO Concentration)- 107.4 µM, 4.4% CV Sample 3 (Nitrite Concentration)- 132.7 µM, 1.3% CV, Sample 1 (Total NO Concentration)- 157.8 µM, 1.8% CV Inter Assay Precision: Three samples were further diluted in Assay Buffer and run in duplicates in twenty assays run over multiple days by three operators. The mean and precision of the calculated Nitrite or Total NO concentrations were: Sample 1 (Nitrite Concentration)- 44.1 µM, 3.1% CV, Sample 1 (Total NO Concentration)- 68.8 µM, 7.4% CV Sample 2 (Nitrite Concentration)- 66.4 µM, 4.0% CV, Sample 1 (Total NO Concentration)- 112.1 µM, 5.7% CV Sample 3 (Nitrite Concentration)- 126.7 µM, 6.3% CV, Sample 1 (Total NO Concentration)- 154.48 µM, 4.1% CV
Incubation Time 30 minutes
Number of Samples 88 samples in duplicate
Other Resources Kit Booklet , MSDS
Field of Use Not for use in humans. Not for use in diagnostics or therapeutics. For in vitro research use only.

Properties

Storage Temperature 4ºC
Shipping Temperature Blue Ice
Product Type Detection Kits
Assay Overview The 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). 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.
Kit Overview
Component No.ItemQuantity / Size
SKC-212A Clear 96 Well Plates 1 Each
SKC-212B Nitrate Standard 200 µl
SKC-212C Nitrite Standard 200 µl
SKC-212D Assay Buffer 60 ml
SKC-212E NADH Concentrate 1.2 ml
SKC-212F Nitrate Reductase 1 Vial
SKC-212G Enzyme Stabilization Buffer 1 ml
SKC-212H Color Reagent A 5 ml
SKC-212I Color Reagent B 5 ml
Cite This Product Nitric Oxide Detection Kit (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SKT-212)

Biological Description

Alternative Names Nitrogen monoxide; Nitrogen(II) oxide Detection Kit
Research Areas Cell Signaling, Neuroscience, Post-translational Modifications
Scientific 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 (1). 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 (2). 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 (3). The balance and dynamics of the regulatory/damage facets of NO are major forces in mitochondrial signaling and dysfunction (4). 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 (5-9).
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) (10).
References 1. Moncada, S and EA Higgs. (1991) Eur. J. Clin. Invest., 21:361-374.
2. Kapil, V. et al. (2010) Heart, 96:1703-1709.
3. Seth, D and Stamler, JS. (2007) Curr. Opin. Chem. Biol., 15:1-8.
4. Eursalimsky, JD and Moncada, S. (2007) ATVB 27:2524-2531.
5. Knott, AB and Bossy-Wetzel, E. (2010) Diab.Obes.Metab. 12(Suppl2):126-133.
6. Van Dyke, K. et al. (2010) Ann. N.Y. Acad. Sci. 1203:138-145.
7. Reher, VSG. et al. (2007) J. Oral Sci. 49(4):271-276.
8. Sogut, S. et al. (2003) Clin. Chim. Acta 331:111-117.
9. Balam. E. et al. (2002) Jpn. J. Clin. Oncol. 32(5):162-166.
10. Moshage, H. (1997) Clin.Chem. 43(4):553-556.

Product Images

<p>Typical Standard Curve for Nitric Oxide Detection Kit StressXpress® – SKT-212. Assay Type: Coupled Enzyme Assay. Detection Method: Colorimetric Assay. Assay Range: 3.125 – 200 uM.</p>

Typical Standard Curve for Nitric Oxide Detection Kit StressXpress® – SKT-212. Assay Type: Coupled Enzyme Assay. Detection Method: Colorimetric Assay. Assay Range: 3.125 – 200 uM.

<p>Linearity was determined by taking two human urine samples with known Nitrite and Total Nitric Oxide Concentrations and mixing them in given ratios. The measured concentrations were compared to the expected values based on the ratios used.</p>

Linearity was determined by taking two human urine samples with known Nitrite and Total Nitric Oxide Concentrations and mixing them in given ratios. The measured concentrations were compared to the expected values based on the ratios used.

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