[KO Validated] Anti-Caspase-8 Rabbit Monoclonal Antibody for WB, ELISA - Q14790

Core Product Specifications and Parameters

Biological Background: Caspase-8 Function and Localization

• Caspase-8 (also designated FLICE, MACH, MCH5) is a 55 kDa initiator caspase of the cysteine-aspartic acid protease family, produced in multiple isoforms through alternative splicing; the full-length isoform 1 is catalytically active while shorter isoforms may act as dominant-negative inhibitors.
• As the key initiator of extrinsic apoptosis, Caspase-8 is recruited via the adaptor FADD to death receptor complexes (DISC) upon FAS or TNFRSF1A stimulation, leading to its dimerization, autocatalytic cleavage, and subsequent activation of downstream effector caspases (CASP3/7) and BID to execute cell death. Related references: PMID:8681376, PMID:8681377, PMID:9184224
• Beyond apoptosis, Caspase-8 negatively regulates necroptosis by cleaving RIPK1 at Asp-324, which inhibits RIPK1 kinase activity and prevents TNF-induced necroptotic and inflammatory signaling. Related references: PMID:31827280, PMID:31827281
• Caspase-8 also mediates pyroptosis by cleaving and activating gasdermin-C (GSDMC) and gasdermin-D (GSDMD), releasing their N-terminal pore-forming domains that cause membrane rupture and inflammatory cell death. Related references: PMID:32929201, PMID:34012073
• It localizes predominantly in the cytoplasm but can be found in the nucleus and at lamellipodia; phosphorylation at Tyr-380, independent of its proteolytic activity, promotes cell migration. Related references: PMID:18216014, PMID:27109099
• Caspase-8 is broadly expressed, with highest mRNA levels in peripheral blood leukocytes, spleen, thymus, and liver, and very low expression in brain, testis, and skeletal muscle.
• Additionally, Caspase-8 cleaves PARP1 and PARP2 during apoptosis, and may participate in Granzyme B-mediated cytotoxic cell death pathways. Related references: PMID:8681376, PMID:8755496

Experimental Guidance and Technical Tips

• The immunogen corresponds to residues 200-300 within the catalytic domain of human Caspase-8; this region is present in the full-length proenzyme and the large catalytic subunit, so the antibody may detect both unprocessed and cleaved forms.
• For Western blot, perform SDS-PAGE under reducing conditions; anticipate a major band at ~55 kDa (full-length) and, depending on cellular treatment, smaller bands representing the active p18/p10 fragments. Always include molecular weight markers.
• This antibody is reportedly KO validated; consider using Caspase-8 knockout cell lysates as a negative control to confirm band specificity in your experimental system.
• For ELISA applications, perform a checkerboard titration to establish optimal coating antigen and antibody concentrations; validate against a known standard if available.
• When studying Caspase-8 activation, treat cells with appropriate inducers (e.g., TNF-α + cycloheximide for apoptosis, or TNF-α + zVAD + Smac mimetic for necroptosis) and monitor cleavage kinetics to distinguish between death modalities.
• Cross-reactivity is reported only for human; if applying to other species, align the immunogenic peptide sequence with the target organism’s Caspase-8 to evaluate potential reactivity.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio specializes in providing diagnostic manufacturers, laboratories, and research institutes with a complete range of IVD raw materials and technical support. This anti-Caspase-8 monoclonal antibody exemplifies our commitment to delivering validated tools for apoptosis and cell death research. Beyond individual antibodies, we offer one-stop access to optimized antibody pairs, bulk ancillary reagents, and rare-target sourcing—helping you bridge the gap from concept to clinical application seamlessly.