Caspase 8 Assay Kit (colorimetric) (ab39700) is a simple and convenient assay to quantify the activity of caspase 8 in cell lysates, based on the recognition of the sequence Ile-Glu-Thr-Asp (IETD).
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- Journal of biochemical and molecular toxicology 36:e231852022Inhibiting AKT signaling pathway with cilostazol and meloxicam synergism for suppressing K562 cells in vitro.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLaya Naderbar,Yaghub Pazhang,Jafar RezaiePubMed 35920412
- Cell death & disease 13:1722022Cell surface nucleolin is a novel ADAMTS5 receptor mediating endothelial cell apoptosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDogan Can Kirman,Bhuvanasundar Renganathan,Wai Kit Chui,Ming Wei Chen,Neslihan Arife Kaya,Ruowen GePubMed 35197459
- Metabolites 11:2021Uncoupling Thermotolerance and Growth Performance in Chinook Salmon: Blood Biochemistry and Immune Capacity.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRonald Lulijwa,Tim Young,Jane E Symonds,Seumas P Walker,Natalí J Delorme,Andrea C AlfaroPubMed 34436488
- International journal of molecular sciences 22:2021Ruthenium(II) and Platinum(II) Complexes with Biologically Active Aminoflavone Ligands Exhibit In Vitro Anticancer Activity.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMałgorzata Fabijańska,Maria M Kasprzak,Justyn OchockiPubMed 34299199
- Turkish journal of biology = Turk biyoloji dergisi 45:171-1792021Positive regulation of TNFR1 signaling via SH3 recognition motif.Applications:Unspecified applicationReactive species:Unspecified reactive speciesFatma Ece Çopuroğlu,Fatma Zehra Hapil,Şükran Burçak Yoldaş,Osman Nidai ÖzeşPubMed 33907493
- Cell metabolism 33:424-436.e102020Apaf-1 Pyroptosome Senses Mitochondrial Permeability Transition.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWanfeng Xu,Yuan Che,Quan Zhang,Hai Huang,Chujie Ding,Yun Wang,Guangji Wang,Lijuan Cao,Haiping HaoPubMed 33308446
- Inorganic chemistry 59:17732-177452020Gold(I) and Silver(I) Complexes with 2-Anilinopyridine-Based Heterocycles as Multitarget Drugs against Colon Cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesInés Mármol,Sara Montanel-Perez,José Carlos Royo,M Concepción Gimeno,M Dolores Villacampa,M Jesús Rodríguez-Yoldi,Elena CerradaPubMed 33205964
- Journal of leukocyte biology 110:343-3562020Inducing regulated necrosis and shifting macrophage polarization with anti-EMMPRIN antibody (161-pAb) and complement factors.Applications:Unspecified applicationReactive species:Unspecified reactive speciesNizar Hijaze,Max Ledersnaider,Elina Simanovich,Sameer Kassem,Michal A RahatPubMed 33205451
- Scientific reports 10:144732020In vitro study on aspects of molecular mechanisms underlying invasive aspergillosis caused by gliotoxin and fumagillin, alone and in combination.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLoganathan Gayathri,Mohammad A Akbarsha,Kandasamy RuckmaniPubMed 32879392
- FASEB journal : official publication of the Federation of American Societies for Experimental Biology 34:13839-138612020Intravitreal administration of adalimumab delays retinal degeneration in rd10 mice.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLorena Olivares-González,Sheyla Velasco,José María Millán,Regina RodrigoPubMed 32816354
Key facts
- Detection method
- Colorimetric
- Sample types
- Cell Lysate
- Assay type
- Enzyme activity
- Assay time
- 2h
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Target data
Function
Thiol protease that plays a key role in programmed cell death by acting as a molecular switch for apoptosis, necroptosis and pyroptosis, and is required to prevent tissue damage during embryonic development and adulthood (PubMed:23516580, PubMed:35338844, PubMed:35446120, PubMed:8681376, PubMed:8681377, PubMed:8962078, PubMed:9006941, PubMed:9184224). Initiator protease that induces extrinsic apoptosis by mediating cleavage and activation of effector caspases responsible for FAS/CD95-mediated and TNFRSF1A-induced cell death (PubMed:23516580, PubMed:35338844, PubMed:35446120, PubMed:8681376, PubMed:8681377, PubMed:8962078, PubMed:9006941, PubMed:9184224). Cleaves and activates effector caspases CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10 (PubMed:16916640, PubMed:8962078, PubMed:9006941). Binding to the adapter molecule FADD recruits it to either receptor FAS/TNFRSF6 or TNFRSF1A (PubMed:8681376, PubMed:8681377). The resulting aggregate called the death-inducing signaling complex (DISC) performs CASP8 proteolytic activation (PubMed:9184224). The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases (PubMed:9184224). Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC (PubMed:9184224). In addition to extrinsic apoptosis, also acts as a negative regulator of necroptosis: acts by cleaving RIPK1 at 'Asp-324', which is crucial to inhibit RIPK1 kinase activity, limiting TNF-induced apoptosis, necroptosis and inflammatory response (PubMed:31827280, PubMed:31827281). Also able to initiate pyroptosis by mediating cleavage and activation of gasdermin-C and -D (GSDMC and GSDMD, respectively): gasdermin cleavage promotes release of the N-terminal moiety that binds to membranes and forms pores, triggering pyroptosis (PubMed:32929201, PubMed:34012073). Initiates pyroptosis following inactivation of MAP3K7/TAK1 (By similarity). Also acts as a regulator of innate immunity by mediating cleavage and inactivation of N4BP1 downstream of TLR3 or TLR4, thereby promoting cytokine production (By similarity). May participate in the Granzyme B (GZMB) cell death pathways (PubMed:8755496). Cleaves PARP1 and PARP2 (PubMed:8681376). Independent of its protease activity, promotes cell migration following phosphorylation at Tyr-380 (PubMed:18216014, PubMed:27109099). Isoform 5. Lacks the catalytic site and may interfere with the pro-apoptotic activity of the complex. Isoform 6. Lacks the catalytic site and may interfere with the pro-apoptotic activity of the complex. Isoform 7. Lacks the catalytic site and may interfere with the pro-apoptotic activity of the complex (Probable). Acts as an inhibitor of the caspase cascade (PubMed:12010809). Isoform 8. Lacks the catalytic site and may interfere with the pro-apoptotic activity of the complex.
Alternative names
MCH5, CASP8, Caspase-8, CASP-8, Apoptotic cysteine protease, Apoptotic protease Mch-5, CAP4, FADD-homologous ICE/ced-3-like protease, FADD-like ICE, ICE-like apoptotic protease 5, MORT1-associated ced-3 homolog, FLICE, MACH
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Recommended products
Caspase 8 Assay Kit (colorimetric) (ab39700) is a simple and convenient assay to quantify the activity of caspase 8 in cell lysates, based on the recognition of the sequence Ile-Glu-Thr-Asp (IETD).
Key facts
- Detection method
- Colorimetric
- Sample types
- Cell Lysate
- Assay type
- Enzyme activity
- Assay time
- 2h
- Assay Platform
- Microplate reader
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
Caspase 8 Assay Kit (colorimetric) (ab39700) is a simple and convenient assay to quantify the activity of caspase 8 in cell lysates, based on the recognition of the sequence Ile-Glu-Thr-Asp (IETD). The assay is based on spectrophotometric detection of the chromophore p-nitroanilide (*p*NA) after it is cleaved from the labeled substrate IETD-*p*NA. The *p*NA light emission can be quantified using a spectrophotometer or a microtiter plate reader at OD 400 – 405 nm. Comparison of the absorbance of *p*NA from an apoptotic sample with an un-induced control allows determination of the fold increase in Caspase 8 activity. Visit our for tips and troubleshooting.
The caspase family of highly conserved cysteine proteases play an essential role in programmed cell death (including apoptosis, pyropoptosis and necroptosis). Mammalian caspases can be subdivided into three functional groups: initiator caspases (Caspase 2, 8, 9 and 10), executioner caspases (Caspase 3, 6 and 7), and inflammatory caspases (Caspase 1, 4, 5, 11 and 12). Initially synthesized as inactive pro-caspases, caspases become rapidly cleaved and activated in response to granzyme B, death receptors and apoptosome stimuli. Caspases will then cleave a range of substrates, including downstream caspases, nuclear proteins, plasma membrane proteins and mitochondrial proteins, ultimately leading to cell death. Caspase 8 (CASP8/FLICE, EC:3.4.22.61) is the most upstream caspase involved in the activation of apoptosis through the extrinsic pathway, mediated by CD95 (Fas) receptor and TNFR. Caspase 8 is recruited to the receptors through the adapter molecule FADD, resulting in the formation of the aggregate called death-inducing signaling complex (DISC) and proteolytic activation of caspase 8. The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases. Inhibition or inactivation of caspase 8 is required for induction of necroptosis. **Other caspase and apoptosis assays** Review the full set of , or the .
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2 product images
Functional Studies - Caspase-8 Assay Kit (Colorimetric) (ab39700)
Active caspase 8 in control (CTRL) Jurkat cells (10e6/mL) or cells treated for five hours with 10 μg/mL Cyclohexamide (CHX) (Cycloheximide, Protein synthesis inhibitor ab120093) or four hours with 25 μg/mL Mitomycin C (MitoC) (Mitomycin C (MMC), Anticancer and antibiotic agent ab120797). Background signal subtracted, duplicates; +/- SD.
Functional Studies - Caspase-8 Assay Kit (Colorimetric) (ab39700)
Active caspase 8 in control (CTRL) Jurkat cells (10e6/mL) or in cells after four hours exposure to 50 ng/mL anti-Fas Ab (αFas) (MBL), or pretreated one hour with 50 μM Z-VAD(OMe)-FMK (Z-VAD(OMe)-FMK, Cell permeable, irreversible pan-caspase inhibitor ab120487) followed by four hours with αFas. Background signal subtracted, duplicates; +/- SD.
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