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Caspase 8 (active) FITC Staining Kit (ab65614) provides a convenient means for sensitive detection of activated caspase 8 in living cells.

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Images

Flow Cytometry - Caspase-8 (active) FITC Staining Kit (AB65614), expandable thumbnail
  • Functional Studies - Caspase-8 (active) FITC Staining Kit (AB65614), expandable thumbnail
  • Functional Studies - Caspase-8 (active) FITC Staining Kit (AB65614), expandable thumbnail

Publications

Key facts

Sample types

Suspension cells, Adherent cells

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 (By similarity). Initiator protease that induces extrinsic apoptosis by mediating cleavage and activation of effector caspases responsible for the TNFRSF6/FAS mediated and TNFRSF1A induced cell death (PubMed:23516580, PubMed:8681376, PubMed:8681377, PubMed:9006941, PubMed:9184224, PubMed:8962078). Cleaves and activates effector caspases CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10 (PubMed:8962078, PubMed:9006941). Binding to the adapter molecule FADD recruits it to either receptor TNFRSF6/FAS mediated or TNFRSF1A (PubMed:8681376, PubMed:8681377). The resulting aggregate called 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-D (GSDMD): GSDMD cleavage promoting release of the N-terminal moiety (Gasdermin-D, N-terminal) that binds to membranes and forms pores, triggering pyroptosis (By similarity). 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 (PubMed:8681376).Isoform 5Lacks the catalytic site and may interfere with the pro-apoptotic activity of the complex.Isoform 6Lacks the catalytic site and may interfere with the pro-apoptotic activity of the complex.Isoform 7Lacks 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 8Lacks the catalytic site and may interfere with the pro-apoptotic activity of the complex.

Alternative names

What's included?

100 Test
Components
FITC-IETD-FMK
1 x 100 µL
Wash Buffer IV
2 x 100 mL
Z-VAD-FMK
1 x 10 µL

Recommended products

Caspase 8 (active) FITC Staining Kit (ab65614) provides a convenient means for sensitive detection of activated caspase 8 in living cells.

Alternative names

Key facts

Sample types

Suspension cells, Adherent cells

Assay type

Enzyme activity

Assay time

2h

Assay Platform

Microplate reader, Fluor. microscope, Flow cyt.

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 (active) FITC Staining Kit (ab65614) provides a convenient means for sensitive detection of activated caspase 8 in living cells. The assay utilizes the caspase 8 inhibitor, IETD-FMK, conjugated to FITC (FITC-IETD-FMK) as a marker. FITC-IETD-FMK is cell permeable, non-toxic, and irreversibly binds to activated caspase 8 in apoptotic cells. The FITC label allows detection of activated caspase-8 in apoptotic cells directly by fluorescence microscopy, flow cytometry, or fluorescence plate reader. Visit our for tips and troubleshooting.

Activation of caspases plays a central role in apoptosis. **Other caspase and apoptosis assays** Review the full set of , or the .

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3 product images

  • Flow Cytometry - Caspase-8 (active) FITC Staining Kit (ab65614), expandable thumbnail
    Hernandez-Flores G et al., BMC Cancer, 11, 483, 2011 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/2.0

    Flow Cytometry - Caspase-8 (active) FITC Staining Kit (ab65614)

    Caspase 8 activation of Hela (left) and SiHa (right) cells after in vitro treatment with pentoxylline (PTX) or cisplatin (CIS) either alone or in combination. Results represent the mean ± SD of three independent experiments carried out in triplicate. (*) p<0.001 vs CTL.
    Image obtained from Hernandez-Flores G et al; BMC Cancer, 2011 Nov 11; 11:483

  • Functional Studies - Caspase-8 (active) FITC Staining Kit (ab65614), expandable thumbnail

    Functional Studies - Caspase-8 (active) FITC Staining Kit (ab65614)

    Active caspase 8 in Jurkat cells following four hours exposure to 50 ng/mL anti-Fas Ab (αFas) (MBL), five hours with 10 μg/mL cyclohexamide (CHX) (Cycloheximide, Protein synthesis inhibitor ab120093), or one hour pretreatment with CHX followed by four hours with αFas. Background signal subtracted, duplicates; +/- SD.

  • Functional Studies - Caspase-8 (active) FITC Staining Kit (ab65614), expandable thumbnail

    Functional Studies - Caspase-8 (active) FITC Staining Kit (ab65614)

    Active caspase 8 in control Jurkat cells (10e6/mL) or cells treated for five hours with 10 ug/mL Cyclohexamide (CHX) (Cycloheximide, Protein synthesis inhibitor ab120093) or four hours with 25 ug/mL Mitomycin C (MitoC) (Mitomycin C (MMC), Anticancer and antibiotic agent ab120797). Background signal subtracted, duplicates; +/- SD.

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