Prostaglandin e2 pge2 competitive elisa kit

The levels of TNF-α (tumor necrosis factor-α) and PGE2 (prostaglandin E2) were measured using Mouse TNF-α ELISA Kit (MULTI SCIENCES, Hangzhou, China) and Prostaglandin E2/PGE2 Competitive ELISA Kit (MULTI SCIENCES, Hangzhou, China), respectively, according to the protocols. In short, 100 μL of diluent standard substance or sample was added onto each well. Then 50 μL diluted antibodies were added and incubated for 2 h at room temperature. After washing, each well was added with 100 μL diluted horseradish peroxidase (HRP)-conjugated streptomycin and incubated for 45 min at room temperature. After repeated washing, 100 μL 3,3ʹ5,5ʹ-Tetramethylbenzidine (TMB) color liquid was added, and the mixture was incubated in the dark at room temperature for 20 min. Finally, 100 μL stop solution was used for stopping the reaction. The optical density (OD) values at the maximum absorption wavelength of 450 nm and the reference wavelength of 570 nm were measured by using a microplate reader (BIOTEK, ELX-800, Vermont, USA). The calibrated OD value was the measured value at 450 nm minus the measured value at 570 nm.