Detect caspase-1 activity with the FAM FLICA Caspase-1 Assay Kit. This in vitro assay employs the fluorescent inhibitor probe FAM-YVAD-FMK to label active caspase-1 enzyme in living cells. Analyze samples using fluorescence microscopy, a fluorescence plate reader, or flow cytometry.

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FAM-FLICA® Caspase-1 (YVAD) Assay Kit
SKU: 97

Size: 25 Tests
Sale price$226.00

Bulk Order FAM-FLICA® Caspase-1 (YVAD) 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-1 probe allows researchers to assess caspase-1 activation. 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 FLICA 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 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.
488 nm / 530 nm
Flow Cytometer, Fluorescence Microscope, Fluorescence Plate Reader
Cell culture
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 97: 25 Tests
  • FLICA Caspase-1 Reagent (FAM-YVAD-FMK), 1 vial, #655
  • 10X Apoptosis Wash Buffer, 15 mL, #635
  • Fixative, 6 mL, #636
  • Propidium Iodide, 1 mL, #638
  • Hoechst 33342, 1 mL, #639
  • Kit Manual
  • Kit 98: 100 Tests
  • FLICA Caspase-1 Reagent (FAM-YVAD-FMK), 4 vials, #655
  • 10X Apoptosis Wash Buffer, 60 mL, #634
  • Fixative, 6 mL, #636
  • Propidium Iodide, 1 mL, #638
  • Hoechst 33342, 1 mL, #639
  • Kit Manual
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    Chen, A;Lu, D;Yang, Z;Che, X;Xia, Y;Shao, X;Chen, Z;Qian, J;Ge, J. Association between NLRP3 inflammasome and periprocedural myocardial injury following elective PCI. Heliyon. 2023 August

    Alam, MA;Caocci, M;Ren, M;Chen, Z;Liu, F;Khatun, MS;Kolls, JK;Qin, X;Burdo, TH. Deficiency of Caspase-1 Attenuates HIV-1-Associated Atherogenesis in Mice. International journal of molecular sciences. 2023 August

    Bourne, JH;Campos, J;Hopkin, SJ;Whitworth, K;Palis, J;Senis, YA;Rayes, J;Iqbal, AJ;Brill, A. Megakaryocyte NLRP3 hyperactivation induces mild anemia and potentiates inflammatory response in mice. Frontiers in immunology. 2023 August

    Wang, D;Wu, Y;Sun, S;Zhao, P;Zhou, X;Liang, C;Ma, Y;Li, S;Zhu, X;Hao, X;Shi, J;Fan, H. NLRP3 inflammasome-mediated pyroptosis involvement in cadmium exposure-induced cognitive deficits via the Sirt3-mtROS axis. The Science of the total environment. 2023 August

    Li, J;Xiao, Y;Zhang, Y;Li, S;Zhao, M;Xia, T;Meng, H. Pulmonary Delivery of Specialized Pro-Resolving Mediators-Based Nanotherapeutics Attenuates Pulmonary Fibrosis in Preclinical Animal Models. ACS Nano. 2023 August

    Wang, D;Yuan, Q;Liu, S;Zhao, P;Liang, C;Ma, Y;Li, S;Zhu, X;Hao, X;Shi, J;Fan, H. BDE-47 flame retardant exposure induces microglial pyroptosis and cognitive deficits by activating the mtROS-NLRP3 axis via Sirt3 downregulation and is salvaged by honokiol. Environmental pollution (Barking, Essex : 1987). 2023 September

    Pampuscenko, K;Morkuniene, R;Krasauskas, L;Smirnovas, V;Brown, GC;Borutaite, V. Extracellular tau stimulates phagocytosis of living neurons by activated microglia via Toll-like 4 receptor-NLRP3 inflammasome-caspase-1 signalling axis. Scientific reports. 2023 July

    Zhang, M;Lan, H;Peng, S;Zhou, W;Wang, X;Jiang, M;Hong, J;Zhang, Q. MiR-223-3p attenuates radiation-induced inflammatory response and inhibits the activation of NLRP3 inflammasome in macrophages. International immunopharmacology. 2023 July

    Kulsuptrakul, J;Turcotte, EA;Emerman, M;Mitchell, PS. A human-specific motif facilitates CARD8 inflammasome activation after HIV-1 infection. eLife. 2023 July

    Xu, K;Qin, X;Zhang, Y;Yang, M;Zheng, H;Li, Y;Yang, X;Xu, Q;Li, Y;Xu, P;Wang, X. Lycium ruthenicum Murr. anthocyanins inhibit hyperproliferation of synovial fibroblasts from rheumatoid patients and the mechanism study powered by network pharmacology. Phytomedicine : international journal of phytotherapy and phytopharmacology. 2023 July

    Kim, JK;Jung, HJ;Hyun, M;Lee, JY;Park, JH;Suh, SI;Baek, WK;Kim, HA. Resistance of hypervirulent Klebsiella pneumoniae to cathepsin B-mediated pyroptosis in murine macrophages. Frontiers in immunology. 2023 June

    Shi, B;Lyu, CJ;Le, ZK;Ji, HS;Xiao, Y;Zhang, YY;Huang, SJ;Yu, LJ;Shu, Q;Tou, JF;Lai, DM. NLRP3 activation in macrophages promotes acute intestinal injury in neonatal necrotizing enterocolitis. World journal of pediatrics : WJP. 2023 June

    Xie, Y;Li, X;Deng, W;Nan, N;Zou, H;Gong, L;Chen, M;Yu, J;Chen, P;Cui, D;Zhang, F. Knockdown of USF2 inhibits pyroptosis of podocytes and attenuates kidney injury in lupus nephritis. Journal of molecular histology. 2023 June

    Miao, RF;Tu, J. LncRNA CDKN2B-AS1 interacts with LIN28B to exacerbate sepsis-induced acute lung injury by inducing HIF-1α/NLRP3-mediated pyroptosis. The Kaohsiung journal of medical sciences. 2023 June

    Li, Z;Shi, Y;Wang, Y;Qi, H;Chen, H;Li, J;Li, L. Cadmium-induced pyroptosis is mediated by PERK/TXNIP/NLRP3 signaling in SH-SY5Y cells. Environmental toxicology. 2023 June

    Liu, L;Feng, L;Gao, J;Hu, J;Li, A;Zhu, Y;Zhang, C;Qiu, B;Shen, Z. Parthenolide targets NLRP3 to treat inflammasome-related diseases. International immunopharmacology. 2023 June

    Zhou, W;Zhang, H;Huang, L;Sun, C;Yue, Y;Cao, X;Jia, H;Wang, C;Gao, Y. Disulfiram with Cu2+ alleviates dextran sulfate sodium-induced ulcerative colitis in mice. Theranostics. 2023 May

    Zhang, Y;Hailati, J;Ma, X;Midilibieke, H;Liu, Z. Ubiquitin-specific protease 11 Aggravates Ischemia-reperfusion-induced Cardiomyocyte Pyroptosis and Injury by Promoting TRAF3 Deubiquitination. Balkan medical journal. 2023 May

    Dong, RJ;Li, J;Zhang, Y;Li, JS;Yang, LH;Kuang, YQ;Wang, RR;Li, YY. Thalidomide promotes NLRP3/caspase-1-mediated pyroptosis of macrophages in Talaromyces marneffei infection. Microbial pathogenesis. 2023 May

    Karema-Jokinen, V;Koskela, A;Hytti, M;Hongisto, H;Viheriälä, T;Liukkonen, M;Torsti, T;Skottman, H;Kauppinen, A;Nymark, S;Kaarniranta, K. Crosstalk of protein clearance, inflammasome, and Ca2+ channels in retinal pigment epithelium derived from age-related macular degeneration patients. The Journal of biological chemistry. 2023 May

    Johnson, AF;Sands, JS;Trivedi, KM;Russell, R;LaRock, DL;LaRock, CN. Constitutive secretion of pro-IL-18 allows keratinocytes to initiate inflammation during bacterial infection. PLoS pathogens. 2023 April

    Li, Q;Hatakeyama, M;Kitaoka, T. Polysaccharide Nanofiber-Stabilized Pickering Emulsion Microparticles Induce Pyroptotic Cell Death in Hepatocytes and Kupffer Cells. Small (Weinheim an der Bergstrasse, Germany). 2023 March

    Brill, A;Bourne, J;Campos, J;Hopkin, S;Whitworth, K;Palis, J;Senis, Y;Rayes, J;Iqbal, A. Megakaryocyte NLRP3 hyperactivation induces anemia and potentiates inflammatory response in mice. Research Square. 2023 March

    Hedbrant, A;Engström, C;Andersson, L;Eklund, D;Westberg, H;Persson, A;Särndahl, E. Occupational quartz and particle exposure affect systemic levels of inflammatory markers related to inflammasome activation and cardiovascular disease. Environmental health : a global access science source. 2023 March

    Sun, L;Li, X;Luo, Z;Li, M;Liu, H;Zhu, Z;Wang, J;Lu, P;Wang, L;Yang, C;Wang, T;He, H;Li, M;Shu, C;Li, J. Purinergic Receptor P2 × 7 Contributes to Abdominal Aortic Aneurysm Development via Modulating Macrophage Pyroptosis and Inflammation. Translational research : the journal of laboratory and clinical medicine. 2023 March

    Cui, Y;Yang, Y;Tao, W;Peng, W;Luo, D;Zhao, N;Li, S;Qian, K;Liu, F. Neutrophil Extracellular Traps Induce Alveolar Macrophage Pyroptosis by Regulating NLRP3 Deubiquitination, Aggravating the Development of Septic Lung Injury. Journal of Inflammation Research. 2023 February

    Ramadan, A;Cao, Z;Gadjeva, M;Zaidi, TS;Rathinam, VA;Panjwani, N. The NLRP3 Inflammasome Is Required for Protection Against Pseudomonas Keratitis. Investigative ophthalmology & visual science. 2023 February

    Lee, H;Do, J;Hong, S;Choi, E;Kim, Y. Potential effects of SORBS1 variants on serum immunoglobulin E levels specific to milk intake in relation to prevalence of atopic dermatitis in Korean children. Journal of Allergy and Clinical Immunology. 2023 February

    Reyes, M;Leff, SM;Gentili, M;Hacohen, N;Blainey, PC. Microscale combinatorial stimulation of human myeloid cells reveals inflammatory priming by viral ligands. Science advances. 2023 February

    Manna, S;Maiti, S;Shen, J;Weiss, A;Mulder, E;Du, W;Esser-Kahn, AP. Nanovaccine that activates the NLRP3 inflammasome enhances tumor specific activation of anti-cancer immunity. Biomaterials. 2023 February

    Chen, Y;Zhou, C;Bian, Y;Fu, F;Zhu, B;Zhao, X;Zhang, M;Zhou, C;Yao, S;Zhang, Z;Luo, H;Ge, Y;Wu, C;Ruan, H. Cadmium exposure promotes thyroid pyroptosis and endocrine dysfunction by inhibiting Nrf2/Keap1 signaling. Ecotoxicology and environmental safety. 2023 January

    Yang, L;Shou, Y;Li, F;Du, J;Zhu, X;Wang, S;Xie, L;Zhou, L;Zhou, J;Yang, Y;Xu, J. The activation of AIM2/Caspase-1/GSDMD pathway contributes to pyroptosis of keratinocytes in Stevens-Johnson syndrome/toxic epidermal necrolysis. Research Square. 2023 January; doi: 10.21203/ PDF

    Zack, S;Nikolaienko, R;Cook, B;Melki, R;Zima, A;Campbell, E. Vacuole Membrane Protein 1 (VMP1) Restricts NLRP3 Inflammasome Activation by Modulating SERCA Activity and Autophagy. Research Square. 2023 January; doi: 10.21203/ PDF

    Zhou, ZY;Bai, SJ;Lu, CW;Yang, B;Wang, J;Lian, S;Jian, ZR;Wang, EL;Wang, GX;Liu, T. Nocardia seriolae mediates liver granulomatous chronic inflammation in Micropterus salmoides through pyroptosis. Journal of fish diseases. 2023 January; doi: 10.1111/jfd.13746. Article

    Hou, L;Zhu, Z;Jiang, F;Zhao, J;Jia, Q;Jiang, Q;Wang, H;Xue, W;Wang, Y;Tian, L. Human umbilical cord mesenchymal stem cell-derived extracellular vesicles alleviated silica induced lung inflammation and fibrosis in mice via circPWWP2A/miR-223-3p/NLRP3 axis. Ecotoxicology and environmental safety. 2023 February; doi: 10.1016/j.ecoenv.2023.114537. Text

    Vaher, H;Kingo, K;Kolberg, P;Pook, M;Raam, L;Laanesoo, A;Remm, A;Tenson, T;Alasoo, K;Mrowietz, U;Weidinger, S;Kingo, K;Rebane, A. Skin colonization with S. aureus can lead to increased NLRP1 inflammasome activation in patients with atopic dermatitis. Journal of Investigative Dermatology. 2023 February; doi: 10.1016/j.jid.2023.01.013. Full Text

    Meyers, AK;Wang, Z;Han, W;Zhao, Q;Zabalawi, M;Duan, L;Liu, J;Zhang, Q;Manne, RK;Lorenzo, F;Quinn, MA;Song, Q;Fan, D;Lin, HK;Furdui, CM;Locasale, JW;McCall, CE;Zhu, X. Pyruvate dehydrogenase kinase supports macrophage NLRP3 inflammasome activation during acute inflammation. Cell reports. 2023 January; doi: 10.1016/j.celrep.2022.111941. Article

    Marques-da-Silva, C;Poudel, B;Baptista, RP;Peissig, K;Hancox, LS;Shiau, JC;Pewe, LL;Shears, MJ;Kanneganti, TD;Sinnis, P;Kyle, DE;Gurung, P;Harty, JT;Kurup, SP. AIM2 sensors mediate immunity to Plasmodium infection in hepatocytes. Proceedings of the National Academy of Sciences of the United States of America. 2023 January; doi: 10.1073/pnas.2210181120. Text

    Mitchell, CA;Verovskaya, EV;Calero-Nieto, FJ;Olson, OC;Swann, JW;Wang, X;Hérault, A;Dellorusso, PV;Zhang, SY;Svendsen, AF;Pietras, EM;Bakker, ST;Ho, TT;Göttgens, B;Passegué, E. Stromal niche inflammation mediated by IL-1 signalling is a targetable driver of haematopoietic ageing. Nature cell biology. 2023 January; doi: 10.1038/s41556-022-01053-0. 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

    Liu, W;Yang, J;Fang, S;Jiao, C;Gao, J;Zhang, A;Wu, T;Tan, R;Xu, Q;Guo, W. Spirodalesol analog 8A inhibits NLRP3 inflammasome activation and attenuates inflammatory disease by directly targeting adaptor protein ASC. The Journal of biological chemistry. 2022 November 12; doi: 10.1016/j.jbc.2022.102696 Research Article

    Liang, C;Peng, Y;Sun, H;Wang, L;Jiang, L;Zou, S. Silencing lncRNA KCNQ1OT1 reduced hepatic ischemia reperfusion injury-induced pyroptosis by regulating miR-142a-3p/HMGB1 axis. Molecular and Cellular Biochemistry. 2022 October 29; doi: 10.1007/s11010-022-04586-y. Full Article

    Jiang, Y;Yang, Y;Hu, Y;Yang, R;Huang, J;Liu, Y;Wu, Y;Li, S;Ma, C;Humphries, F;Wang, B;Wang, X;Hu, Z;Yang, S. Gasdermin D restricts anti-tumor immunity during PD-L1 checkpoint blockade. Cell reports. 2022 October 25; doi: 10.1016/j.celrep.2022.111553. Text

    Moraes, C;Hottz, E;Dos Santos Ornellas, D;Adesse, D;de Azevedo, C;d’Avila, J;Zaverucha-do-Valle, C;Maron-Gutierrez, T;Barbosa, H;Bozza, P;Bozza, F. Microglial NLRP3 Inflammasome Induces Excitatory Synaptic Loss Through IL-1β-Enriched Microvesicle Release: Implications for Sepsis-Associated Encephalopathy. Molecular Neurobiology. 2022 October 25; doi: 10.1007/s12035-022-03067-z. Abstract

    Hu, R;Liang, J;Ding, L;Zhang, W;Liu, X;Song, B;Xu, Y. Edaravone dexborneol provides neuroprotective benefits by suppressing NLRP3 inflammasome-induced microglial pyroptosis in experimental ischemic stroke. International immunopharmacology. 2022 October 21; doi: 10.1016/j.intimp.2022.109315. Article

    Yuan, X;Bhat, OM;Zou, Y;Li, X;Zhang, Y;Li, PL. Endothelial Acid Sphingomyelinase Promotes NLRP3 Inflammasome and Neointima Formation during Hypercholesterolemia. Journal of lipid research. 2022 October 14; doi: 10.1016/j.jlr.2022.100298. Full Text

    Wan, Y;Zhang, W;Huang, C;Jian, J;Zhang, Y;Liu, Q;Chen, P;Zhu, X. Ursolic acid alleviates Kupffer cells pyroptosis in liver fibrosis by the NOX2/NLRP3 inflammasome signaling pathway. International immunopharmacology. 2022 October 14; doi: 10.1016/j.intimp.2022.109321. Text

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    Sun, Y;Han, R;Wang, J;Qin, Y;Ren, Z;Feng, X;Liu, Q;Wang, X. A single-beam of light priming the immune responses and boosting cancer photoimmunotherapy. Journal of Controlled Release : Official Journal of the Controlled Release Society. 2022 September 8; doi: 10.1016/j.jconrel.2022.08.057. Full Article

    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

    Yin, H;Wu, M;Lu, Y;Wu, X;Yu, B;Chen, R;Lu, J;Tong, H. HMGB1-activatied NLRP3 inflammasome induces thrombocytopenia in heatstroke rat. PeerJ. 2022 August 4; doi: 10.7717/peerj.13799. Full Text

    Trugilho, MRO;Azevedo-Quintanilha, IG;Gesto, JSM;Moraes, ECS;Mandacaru, SC;Campos, MM;Oliveira, DM;Dias, SSG;Bastos, VA;Santos, MDM;Carvalho, PC;Valente, RH;Hottz, ED;Bozza, FA;Souza, TML;Perales, J;Bozza, PT. Platelet proteome reveals features of cell death, antiviral response and viral replication in covid-19. Cell death discovery. 2022 July 16; doi: 10.1038/s41420-022-01122-1. Full Text

    McLemore, AF; Hou, HA; Meyer, BS; Lam, NB; Ward, GA; Aldrich, AL; Rodrigues, MA; Vedder, A; Zhang, L; Padron, E; Vincelette, ND; Sallman, DA; Abdel-Wahab, O; List, AF; McGraw, KL. Somatic gene mutations expose cytoplasmic DNA to co-opt the cGAS-STING-NLRP3 axis in Myelodysplastic syndromes. JCI Insight. 2022 July 5; doi: 10.1172/jci.insight.159430. Article

    Panicker, N;Kam, TI;Wang, H;Neifert, S;Chou, SC;Kumar, M;Brahmachari, S;Jhaldiyal, A;Hinkle, JT;Akkentli, F;Mao, X;Xu, E;Karuppagounder, SS;Hsu, ET;Kang, SU;Pletnikova, O;Troncoso, J;Dawson, VL;Dawson, TM. Neuronal NLRP3 is a parkin substrate that drives neurodegeneration in Parkinson's disease. Neuron. 2022 May 25; doi: 10.1016/j.neuron.2022.05.009. Full Article

    Dufies, O;Doye, A;Courjon, J;Torre, C;Michel, G;Loubatier, C;Jacquel, A;Chaintreuil, P;Majoor, A;Guinamard, RR;Gallerand, A;Saavedra, PHV;Verhoeyen, E;Rey, A;Marchetti, S;Ruimy, R;Czerucka, D;Lamkanfi, M;Py, BF;Munro, P;Visvikis, O;Boyer, L. Escherichia coli Rho GTPase-activating toxin CNF1 mediates NLRP3 inflammasome activation via p21-activated kinases-1/2 during bacteraemia in mice. Nature Microbiology. 2021 Jan 11; doi: 10.1038/s41564-020-00832-5. Full Text

    Böhme J, Martinez N, Li S, Lee A, Marzuki M, Tizazu AM, Ackart D, Frenkel JH, Todd A, Lachmandas E, Lum J, Shihui F, Ng TP, Lee B, Larbi A, Netea MG, Basaraba R, van Crevel R, Newell E, Kornfeld H, Singhal A. Metformin enhances anti-mycobacterial responses by educating CD8+ T-cell immunometabolic circuits. Nat Commun. 2020 Oct 16;11(1):5225. doi: 10.1038/s41467-020-19095-z. Full Text

    Gaul S, Leszczynska A, Alegre F, Kaufmann B, Johnson CD, Adams LA, Wree A, Damm G, Seehofer D, Calvente CJ, Povero D, Kisseleva T, Eguchi A, McGeough MD, Hoffman HM, Pelegrin P, Laufs U, Feldstein AE. Hepatocyte pyroptosis and release of inflammasome particles induce stellate cell activation and liver fibrosis. J Hepatol. 2020 Aug 4;S0168-8278(20)30522-5. doi: 10.1016/j.jhep.2020.07.041. Online ahead of print. Abstract

    Chang Y, Zhu J, Wang D, Li H, He Y, Liu K, Wang X, Peng Y, Pan S, Huang K. NLRP3 inflammasome-mediated microglial pyroptosis is critically involved in the development of post-cardiac arrest brain injury. J Neuroinflammation. 2020 Jul 23;17(1):219. doi: 10.1186/s12974-020-01879-1. Full Text

    Huang Y, Wang H, Hao Y, Lin H, Dong M, Ye J, Song L, Wang Y, Li Q, Shan B, Jiang Y, Li H, Shao Z, Kroemer G, Zhang H, Bai L, Jin T, Wang C, Ma Y, Cai Y, Ding C, Liu S, Pan Y, Jiang W, Zhou R. Myeloid PTEN promotes chemotherapy-induced NLRP3-inflammasome activation and antitumour immunity. Nat Cell Biol. 2020 Jun;22(6):716-727. doi: 10.1038/s41556-020-0510-3. Epub 2020 May 4. Abstract

    Hu JJ, Liu X, Xia S, Zhang Z, Zhang Y, Zhao J, Ruan J, Luo X, Lou X, Bai Y, Wang J, Hollingsworth LR, Magupalli VG, Zhao L, Luo HR, Kim J, Lieberman J, Wu H. FDA-approved disulfiram inhibits pyroptosis by blocking gasdermin D pore formation. Nat Immunol. 2020 May 4. doi: 10.1038/s41590-020-0669-6. Online ahead of print. Abstract

    Reis AS, Barboza R, Murillo O, Barateiro A, Peixoto EPM, Lima FA, Gomes VM, Dombrowski JG, Leal VNC, Araujo F, Bandeira CL, Araujo RBD, Neres R, Souza RM, Costa FTM, Pontillo A, Bevilacqua E, Wrenger C, Wunderlich G, Palmisano G, Labriola L, Bortoluci KR, Penha-Gonçalves C, Gonçalves LA, Epiphanio S, Marinho CRF. Inflammasome activation and IL-1 signaling during placental malaria induce poor pregnancy outcomes. Sci Adv. 2020 Mar 4;6(10):eaax6346. doi: 10.1126/sciadv.aax6346. eCollection 2020 Mar. Full Text

    Gao J, Peng S, Shan X, Deng G, Shen L, Sun J, Jiang C, Yang X, Chang Z, Sun X, Feng F, Kong L, Gu Y, Guo W, Xu Q, Sun Y. Inhibition of AIM2 inflammasome-mediated pyroptosis by Andrographolide contributes to amelioration of radiation-induced lung inflammation and fibrosis. Cell Death Dis. 2019 Dec 20;10(12):957. doi: 10.1038/s41419-019-2195-8.>Full Text

    Laencina L, Dubois V, Le Moigne V, Viljoen A, Majlessi L, Pritchard J, Bernut A, Piel L, Roux AL, Gaillard JL, Lombard B, Loew D, Rubin EJ, Brosch R, Kremer L, Herrmann JL, Girard-Misguich F. Identification of genes required for Mycobacterium abscessus growth in vivo with a prominent role of the ESX-4 locus. Proc Natl Acad Sci U S A. 2018. Jan 30;115(5):E1002-E1011. doi: 10.1073/pnas.1713195115. Epub 2018 Jan 17. Abstract

    Mantegazza AR, Wynosky-Dolfi MA, Casson CN, Lefkovith AJ, Shin S, Brodsky IE, Marks MS. Increased autophagic sequestration in adaptor protein-3 deficient dendritic cells limits inflammasome activity and impairs antibacterial immunity. PLoS Pathog. 2017. Dec 18;13(12):e1006785. doi: 10.1371/journal.ppat.1006785. eCollection 2017 Dec. Full text

    Clerc P, Jeanjean P, Hallalli N, Gougeon M, Pipy B, Carrey J, Fourmy D, Gigoux V. Targeted Magnetic Intra-Lysosomal Hyperthermia produces lysosomal reactive oxygen species and causes Caspase-1 dependent cell death. J Control Release. 2017. Dec 1;270:120-134. doi: 10.1016/j.jconrel.2017.11.050. [Epub ahead of print]. Abstract

    Mendonça R, Ferro KP, Leonardo FC, Silva JA, Pericole FV, Saad ST, Costa FF, Conran N. Canonical Inflammasome Formation in Monocytes of Sickle Cell Anemia Patients. Blood. 2017. 130:2233. Abstract

    Bruder-Nascimento T, Ferreira NS, Zanotto CZ, Ramalho F, Pequeno IO, Olivon VC, Neves KB, Alves-Lopes R, Campos E, Silva CA, Fazan R, Carlos D, Mestriner FL, Prado D, Pereira FV, Braga T, Luiz JP, Cau SB, Elias PC, Moreira AC, Câmara NO, Zamboni DS, Alves-Filho JC, Tostes RC. NLRP3 Inflammasome Mediates Aldosterone-Induced Vascular Damage. Circulation. 134(23): 1866-1880. Abstract.

    Zhai Z, Liu W, Kaur M, Luo Y, Domenico J, Samson JM, Shellman YG, Norris DA, Dinarello CA, Spritz RA, Fujita M. NLRP1 promotes tumor growth by enhancing inflammasome activation and suppressing apoptosis in metastatic melanoma. Oncogene. 2017. Abstract.

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    Question: What is the difference between YVAD and WEHD and when is it an advantage using one or the other?

    Answer: Our new product offering FAM-WEHD-FMK is similar to our existing FAM-YVAD-FMK Assay. Both of these peptide sequences are known to target caspase 1,4, and 5. The WEHD sequence is thought to be a “better” caspase-1 target, as the kcat/kM rate is higher for WEHD vs YVAD (meaning faster conversion of substrate product by the enzyme). However, please note that if our understanding of how FLICA works is correct, the FLICA probe never actually binds to the enzyme via the YVAD or WEHD sequence, but rather the FMK moiety, then perhaps these faster binding kinetics are something of a moot point. In practice the performance characteristics of the two product are very similar. In our lab they were shown to be virtually indistinguishable. Nevertheless, we decided to carry both options so that customers can select their preferred targeting sequence based on their individual needs and experience.

    Question: Customer is not seeing a difference between control and induced cells(induction with LPS+ATP). Can we help with optimization? Parameters: macrophages induced from THP-1 cells, using 50 ng/ml PMA for 48 hr Cells in 12 well plates at 3×10^5 cells/well Three groups: experimental with HIV, Positive Control and Untreated. Given fresh media 24 hrs then added 1 ug/ml LPS for 24 hr then 5 mM ATP for 2 hr

    Answer: In our lab, we actually saw a greater response in the THP-1 monocytes (not PMA-primed), we had the greatest response with LPS exposure at 100 ng/mL + 5 mM ATP for 24 hours. In our THP I monocyte studies we found induction levels ranging from 10-30% (average was 26.2%) in 24 hour (LPS/ATP exposure) samples compared to 3-8% in negative controls. When working with THP-1 cells primed with PMA to become macrophage-like, in general we were able to achieve better results with lower LPS concentrations and exposure periods than with the THP-1 monocytes. For instance, exposure to 10ng/mL LPS for 2 hours without any supplemental ATP was sufficient to produce the desired effect. I am a bit concerned that the customer’s use of 1 ug/mL LPS for 24 hours may be too high concentration/exposure period and the susceptible cells are moving through pyroptosis, lysing, and are lost from the positive control sample well prior to even receiving the FAM-YVAD-FMK stain. If this is the case, they are missing the period when more of the positive control cells would be stain positive with FAM-FLICA. I would encourage them to experiment with lower LPS concentrations and exposure periods and see if their results are improved. It is also important to note caspase-1 is rapidly secreted by macrophages after its activation by the inflammasome pathway. Therefore, it turns out macrophages might not be the best cell model for use with this product. We have also been working with nigericin, as an alternate inducing agent.

    Question: The component FAM-YVAD-FMK Part#665 vial in the kit is empty. Please help me to solve this problem.

    Answer: All of our FLICA products, including FAM-YVAD-FMK, are lyophilized as part of the manufacturing process. The vials contain such a small amount of material (µg quantities) that the green FAM-FLICA reagents are nearly invisible in the amber vials. It may be visible as a slight iridescent sheen on the sides of the vial. Per the instructions in our manual, the FLICA vials are reconstituted in DMSO and diluted into PBS and subsequently diluted into cell culture media for staining cells. In order to check that the FLICA vial contains the proper lyophilized reagent, please check the appearance of the DMSO-reconstituted FLICA reagent. It should be orange in appearance and once diluted 1:5 in PBS, the FAM-FLICA reagent should be yellow in appearance.

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