Unicam uv 300

The fruit flies were cultured and treated as above mentioned approach and sacrificed at selected time points (days 10, 20, 30, 40, and 50) (Fig. 1a). The fruit flies (40 flies per sample) were homogenized in 0.40 ml of cold saline followed by centrifugation at a speed of 6,000 g for 10 min at 4°C. Then the supernatant transferred to new centrifugal tubes and was used for each test. SOD activity was quantified using a spectrophotometer (UNICAM, UV-300) according to a SOD assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). This assay is based on the inhibition of nitrite formation from hydroxyl ammonium in the presence of O₂⁻ generators mediated by SOD. The spectrophotometric absorbance was set at 550 nm and the results were expressed as units of SOD activity per mg protein. The protein contents were measured using commercial kit (Nanjing Jiancheng Bioengineering Company) by the coomassie brilliant blue method according to the manufacturer’s directions. Lipid peroxidation products were measured by commercial MDA kit (Nanjing Jiancheng Bioengineering Institute) using a spectrophotometer at 532 nm according to the manufacturer’s directions. The level of lipid peroxidation was calculated and expressed as mmol MDA per mg protein. Every experiment repeated three times.