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.

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Pyroptosis/Caspase-1 Assay, Green
SKU: 9145

Size: 25 - 50 Tests
Price:
Sale price$316
Inflammatory caspases, such as caspase-1, play a critical role in the mechanism to trigger the lytic programmed cell death known as pyroptosis. ICT’s Pyroptosis/Caspase-1 Assay, Green can be used by researchers to assess caspase-1 activity in pyroptotic cells and utilize nigericin as a positive control. The FLICA reagent FAM-YVAD-FMK enters each cell and irreversibly binds to activated caspase-1. Because the FAM-YVAD-FMK FLICA reagent becomes covalently coupled to the active enzyme, it is retained within the cell, while any unbound FAM-YVAD-FMK reagent diffuses out of the cell and is washed away. The remaining green fluorescent signal is a direct measure of the active caspase-1 enzyme activity present in the cell at the time the reagent was added. Cells that contain the bound FLICA can be analyzed by a fluorescent plate reader, fluorescence microscopy, or flow cytometry. Cells labeled with the FLICA reagent may be read immediately or preserved for 24 hours using the fixative. Unfixed samples may also be counter-stained with Propidium Iodide, 7-AAD, or Hoechst (included in the kit). Nigericin induces a net decrease in intracellular levels of potassium, crucial for activation of caspase-1. In pyroptosis experiments, nigericin can be used as a positive control to generate a robust caspase-1 activation response in a variety of cell lines.
FAM-YVAD-FMK
Caspase 1
488 nm / 430 nm
Flow cytometry, Fluorescence microscope, fluorescence plate reader
Cell culture, tissue
Nigericin at ≤ -20°C other components at 2-8°C
Domestic: Overnight Delivery; International: Priority Shipping
United States
  1. Prepare samples and controls.
  2. Dilute 10X Cellular Assay Buffer 1:10 with diH2O.
  3. Reconstitute FAM-FLICA with 50 µL DMSO.
  4. Dilute FAM-FLICA 1:5 by adding 200 µL PBS.
  5. 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).
  6. Incubate approximately 1 hour.
  7. Remove media and wash cells 3 times: add 1X Cellular Wash Buffer and spin cells.
  8. If desired, label with additional stains, such as Hoechst 33342, 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 excited at 492 nm and emits at 520 nm.
Kit 9145: 25-50 Tests
  • FLICA Caspase-1 Inhibitor Reagent (FAM-YVAD-FMK), 1 vial, #655
  • 10X Cellular Wash Buffer, 15 mL, #6164
  • Fixative, 6 mL, #636
  • Hoechst 33342, 1 mL, #639
  • Nigericin, 0.5 µmoles, #6698
  • Kit Manual
  • Kit 9146: 100-200 Tests
  • FLICA Caspase-1 Inhibitor Reagent (FAM-YVAD-FMK), 4 vials, #655
  • 10X Cellular Wash Buffer, 60 mL, #6165
  • Fixative, 6 mL, #636
  • Hoechst 33342, 1 mL, #639
  • Nigericin, 0.5 µmoles, #6698
  • Kit Manual
  • 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

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