- 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.
Gioulbasani M, Galaras A, Grammenoudi S, Moulos P, Dent AL, Sigvardsson M, Hatzis P, Kee BL, Verykokakis M. The transcription factor BCL-6 controls early development of innate-like T cells. Nat Immunol. 2020 Sep;21(9):1058-1069. doi: 10.1038/s41590-020-0737-y. Epub 2020 Jul 27. Abstract
Dodard G, Tata A, Erick TK, Jaime D, Miah SMS, Quatrini L, Escalière B, Ugolini S, Vivier E, Brossay L. Inflammation-Induced Lactate Leads to Rapid Loss of Hepatic Tissue-Resident NK Cells. Cell Rep. 2020 Jul 7;32(1):107855. doi: 10.1016/j.celrep.2020.107855. Full Text
Wandel MP, Kim BH, Park ES, Boyle KB, Nayak K, Lagrange B, Herod A, Henry T, Zilbauer M, Rohde J, MacMicking JD, Randow F. Guanylate-binding proteins convert cytosolic bacteria into caspase-4 signaling platforms. Nat Immunol. 2020 Aug;21(8):880-891. doi: 10.1038/s41590-020-0697-2. Epub 2020 Jun 15. Abstract
Lin JX, Du N, Li P, Kazemian M, Gebregiorgis T, Spolski R, Leonard WJ. Critical functions for STAT5 tetramers in the maturation and survival of natural killer cells. Nat Commun. 2017. Nov 6;8(1):1320. doi: 10.1038/s41467-017-01477-5. Full Text
Chen YL, Xu G, Liang X, Wei J, Luo J, Chen GN, Yan XD, Wen XP, Zhong M, Lv X. Inhibition of hepatic cells pyroptosis attenuates CLP-induced acute liver injury. Am J Transl Res. 2016. Full text.