Griess Reagent Kit [for Nitrite quantitation]

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Supplemental Product Information

Size	Unit Price	Belgium	Japan*	Quantity
1KIT	€240.00	Contact Us	16

*Stock available in Belgium will be delivered in 1 to 3 days
*Stock available in Japan will be delivered in 1 to 2 weeks (excludes regulated items and dry ice shipments).

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Components

• NED Solution (0.1 % N-(1-Naphthyl)ethylenediamine Dihydrochloride) : 100 mL
• Sulfanilamide Solution (1.0 % Sulfanilamide) : 100 mL
• Nitrite Standard (100 mM Sodium nitrite) : 1 mL

Description

• The Griess method is most commonly used to measure the concentration of NO₂^-, a major oxidation product of nitric oxide, in solution. As both NO₂^- and NO₃^- are stable oxidation products of NO, to accurately measure NO levels indirectly from NO₂^-, NO₃^- must be enzymatically reduced to NO₂^- using nitrate reductase (not included in this kit) prior to performing the assay so that the total amount of NO₂^- can be measured.

Storage

• Store at 4 ºC. This product is photosensitive and should be protected from light. Use within 1 year.

Direction for Use

• 1. Bring the Griess Reagent Kit to room temperature. The NED solution may become discolored. However, this discoloration does not significantly affect product performance.
  2. Prepare a 100 µM nitrite solution by diluting the supplied Nitrite Standard (100mM Sodium nitrite) 1:1,000 in the same matrix or buffer used for experimental samples. Perform serial two-fold dilutions of the 100 µM working nitrite standard solution. The final concentrations of nitrite in the standard curve should be as follows: 100, 50, 25, 12.5, 6.25, 3.13, and 1.56 µM. A blank of matrix/buffer should also be prepared. To accurately quantify NO₂^-, construct a calibration curve with a nitrite standard using the same matrix or buffer as the test sample.
  3. Add 50 µl of test sample or nitrite standard to each well of a 96-well plate.
  4. Add 50 µl of Sulfanilamide Solution (1.0% Sulfanilamide) per well. Incubate for 5 minutes at room temperature.
  5. Add 50 µl of NED Solution (0.1% NED) per well.
  6. Incubate for 5-10 minutes at room temperature. The solution will turn a pinkish color in the presence of nitrite.
  7. Measure the absorbance at 540 nm within 30 minutes using a microplate reader. Other wavelengths in the range of 520-560 nm may be used if 540 nm is not available on your instrument.
  8. Plot a standard curve by plotting the corrected blank absorbance at 540 nm for each standard or sample against its concentration in µM. The corrected blank absorbance is calculated by subtracting the average absorbance of the blank solution at 540 nm from that of the standard solution. Using the standard curve, determine the nitrite concentration of each unknown sample.