FAM-FLICA® Caspase-8 Assay Kit
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.
- Prepare samples and controls
- Dilute 10X Apoptosis Wash Buffer 1:10 with diH20.
- Reconstitute FLICA with 50 μL DMSO.
- Dilute FLICA 1:5 by adding 200 μL PBS.
- Add diluted FLICA to each sample at 1:30 (e.g., add 10 μL to 290 μL of cultured cells).
- Incubate approximately 1 hour.
- Remove media and wash cells 3 times: add 1X Apoptosis Wash Buffer and spin cells.
- If desired, label with additional stains, such as Hoechst, Propidium Iodide, 7-AAD, or an antibody.
- If desired, fix cells.
- 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.
Gornowicz, A;Lesyk, R;Czarnomysy, R;Holota, S;Shepeta, Y;Popławska, B;Podolak, M;Szymanowski, W;Bielawski, K;Bielawska, A. Multi-Targeting Anticancer Activity of a New 4-Thiazolidinone Derivative with Anti-HER2 Antibodies in Human AGS Gastric Cancer Cells. International journal of molecular sciences. 2023 April
Peng, Z;Gillissen, B;Richter, A;Sinnberg, T;Schlaak, MS;Eberle, J. Enhanced Apoptosis and Loss of Cell Viability in Melanoma Cells by Combined Inhibition of ERK and Mcl-1 Is Related to Loss of Mitochondrial Membrane Potential, Caspase Activation and Upregulation of Proapoptotic Bcl-2 Proteins. International journal of molecular sciences. 2023 March
Ivasechko, I;Lozynskyi, A;Senkiv, J;Roszczenko, P;Kozak, Y;Finiuk, N;Klyuchivska, O;Kashchak, N;Manko, N;Maslyak, Z;Lesyk, D;Karkhut, A;Polovkovych, S;Czarnomysy, R;Szewczyk, O;Kozytskiy, A;Karpenko, O;Khyluk, D;Gzella, A;Bielawski, K;Bielawska, A;Dzubak, P;Gurska, S;Hajduch, M;Stoika, R;Lesyk, R. Molecular design, synthesis and anticancer activity of new thiopyrano[2,3-d]thiazoles based on 5-hydroxy-1,4-naphthoquinone (juglone). European journal of medicinal chemistry. 2023 March
Rok, J;Rzepka, Z;Banach, K;Kowalska, J;Wrześniok, D. The Assessment of the Phototoxic Action of Chlortetracycline and Doxycycline as a Potential Treatment of Melanotic Melanoma—Biochemical and Molecular Studies on COLO 829 and G-361 Cell Lines. International Journal of Molecular Sciences. 2023 January; doi: 10.3390/ijms24032353. Article
Chao, YY;Puhach, A;Frieser, D;Arunkumar, M;Lehner, L;Seeholzer, T;Garcia-Lopez, A;van der Wal, M;Fibi-Smetana, S;Dietschmann, A;Sommermann, T;Ćiković, T;Taher, L;Gresnigt, MS;Vastert, SJ;van Wijk, F;Panagiotou, G;Krappmann, D;Groß, O;Zielinski, CE. Human TH17 cells engage gasdermin E pores to release IL-1α on NLRP3 inflammasome activation. Nature immunology. 2023 February; doi: 10.1038/s41590-022-01386-w. Article
Janowska, S;Khylyuk, D;Gornowicz, A;Bielawska, A;Janowski, M;Czarnomysy, R;Bielawski, K;Wujec, M. Synthesis and Anticancer Activity of 1,3,4-Thiadiazoles with 3-Methoxyphenyl Substituent. Molecules (Basel, Switzerland). 2022 October 17; doi: 10.3390/molecules27206977. Text
He, L;Wei, T;Huang, Y;Zhang, X;Zhu, D;Liu, H;Wang, Z. miR-214-3p Deficiency Enhances Caspase-1-Dependent Pyroptosis of Microglia in White Matter Injury. Journal of Immunology Research. 2022 August 22; doi: 10.1155/2022/1642896. Full Article
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.