8 isoprostane eliza kit

Eight-isoprostane belongs to F2-isoprostane class. The quantification of 8-isoprostane in urine samples was performed in accordance with the manufacturer’s instructions (8 isoprostane ELIZA kit, Enzo). Briefly, 50 μL of 8-iso-PGF2α conjugate solution, 50 μL of 8-iso-PGF2α antibody solution, and 100 μL of sample were sequentially added to 96-well plates (panel kits), and the plates were incubated 2 h at room temperature on a plate shaker at 500 rpm. The well contents were removed, and the plates were washed three times with 400 μL of wash buffer. In total, 200 mL of the pNpp substrate solution was added to each well, and the plates were incubated for 45 min at room temperature without shaking. Finally, 50 μL of stop solution was added, and the plates were read immediately at 405 nm. The median fluorescence intensities of the targets were analyzed to calculate the corresponding concentrations in the samples. The measurement of urine 8-isoprostane levels was standardized using urinary creatinine levels.

The quantification of 8-OHdG, 8-isoprostane, and TAC in urine samples was performed in accordance with the respective manufacturer’s instructions (8-OHdG ELISA kit, Biovision, Waltham, MA, USA; 8-isoprostane ELIZA kit, Enzo, Farmingdale, NY, USA; Total Antioxidant Capacity Assay Kit, abcam, Cambridge, MA, USA). The laboratory procedures were similar to those reported in a previous study [19 (link)].

The quantifications of 8-OHdG, 8-isoprostane, and total antioxidant capacity (TAC) in urine samples were performed in accordance with the manufacturer’s instructions (8-OHdG ELISA kit, Biovision, Waltham, MA, USA, K4160-100; 8-isoprostane ELIZA kit, Enzo, Farmingdale, NY, USA, DI-900-010; and TAC Assay Kit, Abcam, Cambridge, MA, USA, ab52635). The procedures used in the urine biomarker assay were in accordance with our previous report [25 (link)]. The measurements of urine oxidative stress biomarker levels were further standardized based on urinary creatinine levels measured using a commercial kit (Enzo Life Sciences Inc., Farmingdale, NY, USA, ADI-907-030A).