[18] 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).
Griess reagent 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.
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).