It is based on the principal of inhibition of NO radical (which is generated from sodium nitroprusside in phosphate buffered saline with the addition of Griess reagent
In this experiment, NO concentration in plasma was measured by Griess reagent
(Promega, USA) system.
An aliquot of the solution of each sample was removed and diluted with Griess reagent
comprising of 1% sulfanilamide in 5% H3PO4, and 0.1% naphthylethylenediamine dihydrochloride.
Nitric oxide production was determined by using Griess reagent
kit (Molecular Probes Life technologies).
kit, Dulbecco's modified eagle medium (DMEM), and fetal calf serum (FCS) were purchased from GIBCO/Life Technologies Inc.
Dulbecco's Modified Eagle's medium (DMEM), 3-(4,5-dimethylthiazol2-yl)-2,5- diphenyltetrazolium bromide (MTT), lipopolysaccharide (LPS, Escherichia coli serotype 055: B5), the Griess reagent
, and the primers were acquired from Sigma Chemical Co.
The supernatants were collected and assessed for nitrite production using Griess reagent
Culture media, isolated, and the concentration of nitrite, an indicator of nitric oxide (NO) synthesis, were measured by using Griess reagent
(1% sulfanilic acid in 5% phosphoric acid and 0.1% N-ethylenediaminedihydrochloride; Molecular Probes, Waltham, MA, USA).
Samples were added to the wells, followed by the immediate addition of Griess reagent
. Griess reagent
was produced by adding equivalent parts of solution 1 containing 1% sulphanilamide diluted in 5% phosphoric acid and solution 2 containing 0.1% N(1-Naphthyl)ethylenediamine dihydrochloride diluted in 5% phosphoric acid.
On 10th day, 0.5 ml of Griess reagent
was added to one drug-free control tube to observe any color change (strong or weak pink).
On the tenth day, 0.5 mL of Griess reagent
was added to one drug free control tube to observe any colour change (Strong or weak pink).