Pyroptosis/Caspase-1 Assay - Far Red Fluorescent

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 (e.g. add 10 μL to 290 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, Propidium Iodide, 7-AAD, or an antibody. 9. If desired, fix cells. 10. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. FAM-FLICA excites at 492 nm and emits at 520 nm. If working with adherent cells, please see the manual for additional protocols.