Pyroptosis/Caspase-1 Assay, Green
- Prepare samples and controls.
- Dilute 10X Cellular Assay Buffer 1:10 with diH2O.
- Reconstitute FAM-FLICA with 50 µL DMSO.
- Dilute FAM-FLICA 1:5 by adding 200 µL PBS.
- Add diluted FAM-FLICA to each sample at 1:30-1:60 (e.g., spike at 1:30 by adding 10 µL to 290 µL sample).
- Incubate approximately 1 hour.
- Remove media and wash cells 3 times: add 1X Cellular Wash Buffer and spin cells.
- If desired, label with additional stains, such as Hoechst 33342, Propidium Iodide, 7-AAD, or an antibody.
- If desired, fix cells.
- Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. FAM-FLICA excited at 492 nm and emits at 520 nm.
Chen, A;Chen, Z;Zhou, Y;Wu, Y;Xia, Y;Lu, D;Fan, M;Li, S;Chen, J;Sun, A;Zou, Y;Qian, J;Ge, J. Rosuvastatin protects against coronary microembolization-induced cardiac injury via inhibiting NLRP3 inflammasome activation. Cell Death & Disease. 2021 Jan 12; 12(1)78. doi: 10.1038/s41419-021-03389-1. Full Article