ICT’s Pyroptosis/Caspase-1 Assay, Far Red utilizes our popular FLICA technology to measure caspase-1 activation. Using this kit, researchers can easily detect pyroptotic cells in their whole cell samples in vitro. This complete kit includes Nigericin for the simple and reliable generation of a positive control. Analyze the fluorescence signal using flow cytometry or fluorescence microscopy.
- Prepare samples and controls.
- Dilute 10X Cellular Assay Buffer 1:10 with diH2O.
- Reconstitute FLICA with 50 µL DMSO.
- Dilute FLICA 1:5 by adding 200 µL PBS.
- Add diluted 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.
- Resuspend cell pellet in 1X Cellular Wash Buffer.
- If desired, label with additional stains, such as Hoechst 33342, DAPI, or an antibody.
- If desired, fix cells.
- Analyze with a fluorescence microscope or flow cytometer. FLICA 660 is excited at 660 nm and emits at 680-690 nm.
Mao, x;Wu, W;Nan, Y;Sun, W;Wang, Y. SMAD2 inhibites pyroptosis of fibroblast-like synoviocytes and secretion of inflammatory factors via TGF-β pathway in rheumatoid arthritis. Research Square. 2023 January; doi: 10.21203/rs.3.rs-2471290/v1. PDF