ICT’s Pyroptosis/Caspase-1 Assay Kit utilizes our popular FLICA technology to detect caspase-1 activation. Using this kit, researchers can easily assess pyroptotic cells and utilize nigericin as a positive control. Analyze the green fluorescent signal using fluorescence microscopy, a fluorescent plate reader, or by flow cytometry.
- 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.
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