Green Fluorescent FAM-VAD-OPH II in vitro poly Caspase Apoptosis Detection Reagent

1. For in vitro apoptosis detection, culture your cells to a concentration of 2-5 x 105 cells/mL. 2. Prepare experimental and control populations; concentrate cells to 5 x 105-106 cells/mL in 300 μL samples. 3. Reconstitute the FAM-OPH reagent with 50 μL DMSO to form the 150X stock concentrate (may be frozen for future use) . 4. Dilute the FAM-OPH stock concentrate with 200 μL PBS to form the 30X working solution. 5. Add FAM-OPH working solution directly to samples and controls at a ratio of 1:30. For example, add 10 μL of working solution to a 300 μL aliquot of suspension cells. 6. Incubate ~1 hour. 7. Wash and spin cells two or three times. 8. If desired, label cells with Hoechst 33342 nuclear stain, DAPI nuclear stain, or other compatible fluorescent markers. 9. If desired, fix cells with formalin-based agents. 10. Analyze data using a fluorescence microscope, plate reader, or flow cytometer.