Caspase 8 Monoclonal Antibody (1H10), eBioscience™, Invitrogen™

Description

• Description: The monoclonal antibody 1H10 recognizes human Caspase-8 also known as FLICE and MACH, a member of the cysteine-requiring aspartate protease family
• Caspase-8 is an upstream caspase responsible for Fas and TNF-alpha mediated apoptosis
• After activation of the Fas or TNF receptors by their ligands, the adaptor molecule FADD binds to the receptor and recruits caspase-8 to the membrane
• The formation of this complex induces the cleavage and activation of caspase-8
• The active form of caspase-8, made of a heterotetramer of p18 and p10, is released in the cytoplasm and activates downstream apoptotic proteases (caspase-3, -4, -6, -7, -9 and -10)
• Several (> 8) isoforms have been identified: 3 different pro-forms called Caspase-8a (55kDA), Caspase-8b (53kDa), and Caspase-8L which is procaspase-8 with an additional N-terminal extension of 59 aa
• The 1H10 antibody recognizes the p55 and p53 proforms in addition to the p43 and p18 cleaved forms
• Under reducing conditions, an additional band (possibly non-specific) around 35kDa may be present
• Under nonreducing conditions, this band is absent
• Applications Reported: This 1H10 antibody has been reported for use in immunoblotting (WB)
• Applications Tested: This 1H10 antibody has been tested by immunoblot analysis of cell lines
• This can be used at 1-5 μg/mL It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest
• Caspase 8 (CASP8) is a member of the cysteine-aspartic acid protease (caspase) family
• Sequential activation of caspases play a central role in the execution-phase of cell apoptosis
• Caspases exist as inactive proenzymes composed of a pro-domain, a large protease subunit, and a small protease subunit
• Activation of caspases requires proteolytic processing at conserved internal aspartic residues to generate a heterodimeric enzyme consisting of the large and small subunits
• Caspase 8 is involved in the programmed cell death induced by Fas and various apoptotic stimuli
• The N-terminal FADD-like death effector domain of Caspase 8 suggests that it may interact with Fas-interacting protein FADD
• Caspase 8 was detected in the insoluble fraction of the affected brain region from Huntington disease patients but not in those from normal controls, which implicated the role in neurodegenerative diseases
• Caspase 8 binds to the death effector domain (DED) of FADD through an analogous DED domain present in tandem in the pro-form of the Caspase 8 protein
• Activated Caspase 8 then activates other downstream caspases including Caspase 9, thereby committing the cell to undergo apoptosis
• In addition, Caspase 8 also reacts with Jurkat cells and Tonsil
• Overexpression of Caspase 8 induces apoptosis, which can be blocked by inhibitors specific for the ICE family
• Many alternatively spliced transcript variants encoding different isoforms have been described for Caspase 8, however, not all variants have had their full-length sequences determined.