4.4. Lipid Peroxidation Assay

About 5 mg of wet retina were collected, washed with saline solution, and homogenized in 300 microliters of RIPA lysis buffer for two min. The thiobarbituric acid reactive substance (TBARS) assay was used, according to the manufacturer’s instructions (OXItek-TBARS assay kit, Enzo Life Sciences, Farmingdale, NY, USA) to quantify of malondialdehyde (MDA) production during acid hydrolysis of the lipid peroxides. Briefly, the mixture was incubated at 95 °C for 1 h and cooled on ice for 10 min. Then, samples were centrifuged at 3000 rpm at 4 °C for 15 min, and the supernatant was used to measure MDA concentration (nmol/mg of protein) at 532 nm (EON, BioTek Instruments, Inc., Winooski, VT, USA). The concentration of MDA was calculated according to the average absorbance of the MDA from a standard curve.