Necrosis vs Apoptosis Assay Kit

1. Prepare samples and controls. 2. Dilute 10X Apoptosis Wash Buffer 1:10 with diH20. 3. Reconstitute FLICA with 50 μL DMSO. 4. Dilute FLICA 1:5 by adding 200 μL PBS. 5. Add diluted FLICA to each sample at 1:30 – 1:60. For example, to stain at 1:30, add 10 μL to 290 μL of cultured cells. To stain at 1:60, add 5 μL to 295 μL of cultured cells. 6. Incubate approximately 1 hour. 7. Remove media and wash cells 3 times: add 1X Apoptosis Wash Buffer and spin cells. 8. If desired, label with additional stains, such as Hoechst, 7-AAD, or an antibody. 9. If desired, fix or embed cells. 10. Analyze with a fluorescence microscope, fluorescence platereader, or flow cytometer. SR-FLICA excites at 550-580 nm and emits at 590-600 nm.