Swine Annexin V-Fluorescein Apoptosis Assay Kit

1. Prepare the 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. Dilute 10X Wash Buffer 1:10 with diH2O., 6. Wash plate 4 times with 1X Wash Buffer., 7. Add 100 µL TMB Substrate to each well., 8. Cover plate with Plate Cover and incubate at room temperature in the dark for 30 minutes to develop the signal., 9. Add 100 µL Stop Solution to each well., 10. Measure absorbance using a plate reader at 450 nm., 11. Plot the standard curve and calculate sample concentrations.