IndexThe human red blood cell (HRBC) membrane stabilization methodInhibition of protein denaturationAnti-proteinase actionAnti-lipoxygenase activityThe human red blood cell (HRBC) membrane stabilization methodBlood was collected from volunteers healthy humans and an equal volume of Alsever solution (2% dextrose, 0.8% sodium citrate, 0.5% citric acid, and 0.42% NaCl) was mixed with it and centrifuged at 3,000 rpm for 10 minutes. The resulting packed cells were washed with normal saline, and a 10% HRBC suspension was prepared. Various concentrations of AOAgNPs (50,100,150,200 and 500 µg ml-1) were prepared using distilled water. A mixture of 1 ml phosphate buffer, 2 ml hyposaline, and 0.5 ml HRBC suspension (of the above various concentrations) was prepared. It was incubated for 30 minutes at 37°C and centrifuged at 3,000 rpm for 20 minutes. The absorbance of the supernatant solution was measured spectrophotometrically at 560 nm (Kamalutheen et al., 2009). Diclofenac sodium was taken as standard medication. The experiment was repeated three times. Say no to plagiarism. Get a tailor-made essay on "Why Violent Video Games Shouldn't Be Banned"? Get an original assayInhibition of protein denaturationTo study the inhibitory effect on protein denaturation, AOAgNPs were added to a 1% aqueous solution of BSA and the pH of the reaction mixture was adjusted. Sample mixtures were incubated at 37°C for 20 minutes and then reheated to 51°C for 20 minutes. After cooling, the turbidity of the samples was measured spectrophotometrically at 660 nm (Deshpande et al., 2009). Diclofenac sodium was used as the standard drug. The experiment was conducted in triplicate. Antiproteinase action The antiproteinase activity of AOAgNPs was estimated according to the modified method of Sakat et al. 2010. Initially, 2 ml of reaction mixture containing 0.06 mg/ml of trypsin, 1 ml of 20 mM Tris HCl buffer (pH 7.4) and 1 ml of AOAgNPs at different concentrations (100 - 500 µg ml) were prepared -1). The mixture was incubated at 37°C for 5 minutes and then 1 ml of 0.8% (w/v) casein was added to it and incubated for 20 minutes. Subsequently, 2 ml of 70% perchloric acid were added to stop the reaction. Subsequently, the entire suspension was centrifuged and the absorbance was measured at 210 nm. Indomethacin was used as the standard drug. Percent inhibition of proteinase inhibitory activity was calculated using the following formula. Please note: this is just a sample. Get a custom paper from our expert writers now. Get a Custom Assay Anti-lipoxygenase activity The anti-lipoxygenase activity of AOAgNPs was studied using linoleic acid as a substrate and lipoxidase as an enzyme (Shinde et al. 1999). Samples were dissolved in 250 µl of 2M borate buffer (pH 9.0) and 250 µl of lipoxidase enzyme solution (20,000 U/ml) and incubated for 5 minutes at 25°C. Next, 1.0 ml of lenoleic acid solution (0.6 mM) was added, mixed well, and the absorbance was measured at 234 nm. Indomethacin was used as the standard drug, and the percentage of inhibition was calculated from the above-mentioned formula.