Nitrotyrosine ELISA Kit

1. Prepare standard and samples in the Sample and Standard Diluent. 2. Add 50 μL of prepared standards and samples in triplicate to appropriate wells. 3. Add 50 μL of the diluted antibody preparation to the appropriate wells. 4. Cover plate with Plate Cover and incubate at room temperature (20-25°C) for 1 hour. 5. Wash plate 4 times with 1X Wash Buffer. 6. Add 100 μL of TMB Substrate to each well. 7. Cover plate and develop the plate in the dark at room temperature for 30 minutes. 8. Add 100 μL of Stop Solution to each well. 9. Measure absorbance on a plate reader at 450 nm. 10. Plot the standard curve and calculate sample concentrations.