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Each lot of this antibody is quality control tested by Western blotting. Western blotting, suggested working dilution(s): Use 5 μg antibody per 5 ml antibody dilution buffer for each mini-gel. It is recommended that the reagent be titrated for optimal performance for each application.
COA:
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Application References:
1. Liu G, et al. 2010. J. Natl Cancer Inst. 102:812. PubMed
Jurkat cell extract was resolved by electrophoresis, transferred to nitrocellulose and probed with monoclonal anti-caspase 8 antibody. Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence system.
Caspase 8 (also known as ICE-like apoptotic protease 5, MORT1-associated CED-3 homolog (MACH), FADD-like ICE (FLICE), and apoptotic protease Mch-5) is a member of the peptidase family C14 that contains a death effector domain. Nine isoforms of this caspase have been reported, pro-Caspase 8 has a molecular weight of 55 kD. This caspase is found in the cytoplasm and, upon activation, in the mitochondria. Caspase 8 is the most upstream protease of caspase activation cascade, responsible for TNFRSF6/FAS/TRAIL-R mediated and TNFRSF1A induced cell death. This caspase cleaves/activates Caspase 3, Caspase 4, Caspase 6, Caspase 7, Caspase 9, Caspase 10, and Bid. Caspase 8 can be inhibited by the dominant negative isoform, ARC, and c-FLIP. Caspase 8 autocleaves into large p18 and small p10 subunits when activated by recruitment to the DISC complex at plasma membrane by FADD. This caspase interacts with FADD, CFLAR, and PEA15 proteins. Caspase 8 isoform 9 interacts at the ER with a complex containing BCAP31, BAP29, Bcl-2 and/or Bcl-2L1. Substrates for caspase 8 include androgen receptor, β-catenin, gelsolin, and vimentin. The 4-1-20 monoclonal antibody has been shown to be useful for Western blotting of human caspase 8 (55 kD pro-caspase form).
Peptidase family C14, death effector domain, heterotetramer. Nine isoforms, pro-Casp8 55 kD
Distribution:
Cytoplasm, when activated mitochondrial
Function:
Most upstream protease of caspase activation cascade, responsible for TNFRSF6/FAS/TRAIL-R mediated and TNFRSF1A induced cell death. Cleaves/activates Casp3, Casp4, Casp6, Casp7, Casp9, Casp10, Bid
Regulation:
Inhibited by dominant negative isoform, ARC, c-FLIP. Autocleaves into large p18 and small p10 subunits when activated by recruitment to DISC at plasma membrane by FADD
Interaction:
FADD, CFLAR, PEA15; isoform 9 interacts at the ER with a complex containing BCAP31, BAP29, Bcl-2 and/or Bcl-2L1. Cleaves androgen receptor, beta-catenin, gelsolin, vimentin
Antigen References:
1. Boldin M, et al. 1996. Cell. 85:803. 2. Breckenridge D, et al. 2002. P. Natl. Acad. Sci. USA 99:4331. 3. Himeji D, et al. 2002. Blood. 99:4070. 4. van Noesel M. 2003. Cancer Lett. 197:165.
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Purified anti-Caspase-8
Jurkat cell extract was resolved by electrophoresis, transferred to nitrocellulose and probed with monoclonal anti-caspase 8 antibody. Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence system.