NO production was measured from cell supernatants of suppression assays (described above) after 65-72 hours of culture, using the Griess reagent for nitrite (Sigma-Aldrich; St. Louis, MO), as previously described (O'Connor, MA et al., in revision).
Griess reagent for nitrite
Manufactured by Merck Group
Sourced in Germany, United States
The Griess reagent for nitrite is a laboratory product that is used to detect and quantify the presence of nitrite ions in various samples. It is a colorimetric reagent that reacts with nitrite to produce a purple-colored azo dye, which can be measured using a spectrophotometer. The Griess reagent is a commonly used tool in analytical chemistry and biochemistry for the analysis of nitrite levels in environmental, biological, and food samples.
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Lab products found in correlation
Nitric oxide, by Merck Group (1 mentions)
Tristar lb 941, by Berthold Technologies (1 mentions)
Gentamycin, by Merck Group (1 mentions)
Trypan blue, by Merck Group (1 mentions)
Tert butyl hydroperoxide, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Gallic acid, by Merck Group (1 mentions)
Histopaque 1077, by Merck Group (1 mentions)
Phosphate buffer saline (pbs), by Merck Group (1 mentions)
Sodium nitroprusside, by Merck Group (1 mentions)
5 protocols using griess reagent for nitrite
1
Nitrite Production Measurement Protocol
2
Evaluating Anti-Inflammatory Potential
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The chemicals used were obtained from the following suppliers: Indomethacin and MTT (Sigma), quercetin, RPMI 1640 bottle (Sigma/Lonza/Highveld Biological), Penicillin/Streptomycin/Fongizone (PSF) (Highveld Biological), BS (Highveld Biological), U937 Cells (ATCC® CRL1593.2 ™), Griess reagent for nitrite (Lot BCBJ6549) Sigma, Germany; Nitric oxide (Lot S38919-456) Sigma, USA; lipopolysaccharide (LPS) from Escherica coli (L6529-1MG) Sigma, Germany;
3
Quantifying Nitric Oxide Production
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Nitric oxide (NO) production was quantified as nitrite accumulation using the Griess reagent for nitrite (Sigma-Aldrich) as described previously [15 (link)]. Briefly, cells were cultured in 48-well plates at a density of 2 × 105 cells/well and treated with aleglitazar (250, 125, 62.5 or 31.3 μM) or vehicle (DMSO) for 48 h. Aliquots of 50 μl cell culture supernatant were incubated with 50 μl Griess’ reagent. Absorption was measured at 550 nm with a microplate spectrophotometer (TriStar LB941, Berthold Technologies, Bad Wildbad, Germany).
4
Antioxidant and Cytotoxicity Assays
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Gallic acid, sodium nitroprusside, Griess reagent for nitrite, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide, RPMI 1640, tert-butyl hydroperoxide, Histopaque-1077, trypan blue, phosphate buffer saline, gentamycin, and fetal bovine serum were purchased from Sigma-Aldrich (St. Louis, MO, USA). Hydrogen peroxide, ascorbic acid, and ethanol were supplied by Labosi (Paris, France).
5
Nitric Oxide Quantification in LPS-Treated Cells
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The cells were grown to confluence in a 6-well plate. Half of the wells were then treated with 100 ng mL -1 lipopolysaccharide (LPS, from salmonella), and arginine monohydrochloride was added to all wells (5 mM final concentration) to give a high concentration of substrate for nitric oxide synthases. After 24 h incubation, the cell culture medium was recovered, centrifuged at 10 000g for 10 minutes to remove cells and debris, and the nitrite concentration was measured after adding an equal volume of Griess reagent (Griess' reagent for nitrite, catalog number 03553 from Sigma-Aldrich). It was then incubated at room temperature for 30 minutes. Absorbance was measured at 540 nm.
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