Detect caspase-8 activity with the FLICA Caspase-8 Assay Kit. This in vitro assay employs the fluorescent inhibitor probe FAM-LETD-FMK to label active caspase-8 enzyme in living cells. Analyze samples using fluorescence microscopy, a fluorescence plate reader, or flow cytometry.

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FAM-FLICA® Caspase-8 Assay Kit
SKU: 99

Size: 25 Tests
Price:
Sale price$201


Bulk Order FAM-FLICA® Caspase-8 Assay Kit

Caspases play important roles in apoptosis and inflammation. ICT’s FLICA assay kits are used by researchers seeking to quantitate apoptosis via caspase activity in cultured cells and tissues. The FAM FLICA Caspase-8 assay probe allows researchers to assess caspase-8 activation. The FLICA reagent FAM-LETD-FMK enters each cell and irreversibly binds to activated caspase-8. Because the FAM-LETD-FMK FLICA reagent becomes covalently coupled to the active enzyme, it is retained within the cell, while any unbound FAM-LETD-FMK FLICA reagent diffuses out of the cell and is washed away. The remaining green fluorescent signal is a direct measure of the active caspase-8 enzyme activity present in the cell at the time the reagent was added. Cells that contain the bound FLICA can be analyzed by a fluorescence plate reader, fluorescence microscopy, or flow cytometry. Cells labeled with the FLICA reagent may be read immediately or preserved for 16 hours using the fixative included in the kit. Unfixed samples may also be analyzed with Propidium Iodide or Hoechst 33342 to detect necrosis or changes in nuclear morphology, respectively.
FAM-LETD-FMK
Caspase-8
488 nm / 530 nm
Flow Cytometer, Fluorescence Microscope, Fluorescence Plate Reader
Cell culture
2-8°C
Domestic: Overnight Delivery; International: Priority Shipping
United States
  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.

Kit 99: 25 Tests
  • FLICA Caspase-8 Reagent (FAM-LETD-FMK), 1 vial, #656
  • 10X Apoptosis Wash Buffer, 15 mL, #635
  • Fixative, 6 mL, #636
  • Propidium Iodide, 1 mL, #638
  • Hoechst 33342, 1 mL, #639
  • Kit Manual
  • Kit 910: 100 Tests
  • FLICA Caspase-8 Reagent (FAM-LETD-FMK), 4 vials, #656
  • 10X Apoptosis Wash Buffer, 60 mL, #635
  • Fixative, 6 mL, #636
  • Propidium Iodide, 1 mL, #638
  • Hoechst 33342, 1 mL, #639
  • Kit Manual
  • Kubiak, AB;Ziółkowska, EI;Korycka-Wołowiec, AB;Robak, T;Wołowiec, D. The influence of venetoclax, used alone or in combination with cladribine (2-CdA), on CLL cells apoptosis in vitro: Preliminary results. Advances in Clinical and Experimental Medicine : Official Organ Wroclaw Medical University. 2022 April 25; doi: 10.17219/acem/148142. Abstract

    Lima, TS;Mallya, S;Jankeel, A;Messaoudi, I;Lodoen, MB. Toxoplasma gondii Extends the Life Span of Infected Human Neutrophils by Inducing Cytosolic PCNA and Blocking Activation of Apoptotic Caspases. mBio. 2021 Jan 26; 12(1). doi: 10.1128/mBio.02031-20. Full Article

    Suzuki T, Kohyama K, Moriyama K, Ozaki M, Hasegawa S, Ueno T, Saitoe M, Morio T, Hayashi M, Sakuma H. Extracellular ADP augments microglial inflammasome and NF-κB activation via the P2Y12 receptor. Eur J Immunol. 2019 Sep 24. doi: 10.1002/eji.201848013. [Epub ahead of print]. https://onlinelibrary.wiley.com/doi/abs/10.1002/eji.201848013. Abstract

    Gornowicz A, Pawłowska N, Czajkowska A, Czarnomysy R, Bielawska A, Bielawski K, Michalak O, Staszewska-Krajewska O, and Kałuża Z. Biological evaluation of octahydropyrazin[2,1-a:5,4-a’]diisoquinoline derivatives as potent anticancer agents. Tumour Biol. 2017. Jun;39(6):1010428317701641. doi: 10.1177/1010428317701641. Full Text.

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