The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Specific Activity: 5000 units/mg. One unit of the recombinant Caspase-8 is the enzyme activity that cleaves 1 nmol of the caspase substrate IETD-pNA (pNA: pnitroanaline) per hour at 37°C in a reaction solution containing 50 mM Hepes, pH 7.2, 50 mM NaCl, 0.1% Chaps, 10 mM EDTA, 5% Glycerol, and 10 mM DTT.
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Preparation and Storage
Stability and Storage
Shipped at 4°C. Upon delivery aliquot. Store at -80°C. Avoid freeze / thaw cycle.
This product is an active protein and may elicit a biological response in vivo, handle with caution.
ReconstitutionReconstitute to 1 unit per µl in PBS containing 15% glycerol.
Amyotrophic lateral sclerosis 2 chromosomal region candidate gene 12 protein
Apoptotic cysteine protease
Apoptotic protease Mch-5
Apoptotic protease Mch5
Caspase 8 apoptosis related cysteine peptidase
Caspase-8 subunit p10
FADD Like ICE
FADD-homologous ICE/CED-3-like protease
ICE-like apoptotic protease 5
MACH alpha 1/2/3 protein
MACH beta 1/2/3/4 protein
MORT1 associated ced 3 homolog
MORT1-associated CED-3 homolog
FunctionMost upstream protease of the activation cascade of caspases responsible for the TNFRSF6/FAS mediated and TNFRSF1A induced cell death. Binding to the adapter molecule FADD recruits it to either receptor. The resulting aggregate called death-inducing signaling complex (DISC) performs CASP8 proteolytic activation. The active dimeric enzyme is then liberated from the DISC and free to activate downstream apoptotic proteases. Proteolytic fragments of the N-terminal propeptide (termed CAP3, CAP5 and CAP6) are likely retained in the DISC. Cleaves and activates CASP3, CASP4, CASP6, CASP7, CASP9 and CASP10. May participate in the GZMB apoptotic pathways. Cleaves ADPRT. Hydrolyzes the small-molecule substrate, Ac-Asp-Glu-Val-Asp- -AMC. Likely target for the cowpox virus CRMA death inhibitory protein. Isoform 5, isoform 6, isoform 7 and isoform 8 lack the catalytic site and may interfere with the pro-apoptotic activity of the complex.
Tissue specificityIsoform 1, isoform 5 and isoform 7 are expressed in a wide variety of tissues. Highest expression in peripheral blood leukocytes, spleen, thymus and liver. Barely detectable in brain, testis and skeletal muscle.
Involvement in diseaseDefects in CASP8 are the cause of caspase-8 deficiency (CASP8D) [MIM:607271]. CASP8D is a disorder resembling autoimmune lymphoproliferative syndrome (ALPS). It is characterized by lymphadenopathy, splenomegaly, and defective CD95-induced apoptosis of peripheral blood lymphocytes (PBLs). It leads to defects in activation of T-lymphocytes, B-lymphocytes, and natural killer cells leading to immunodeficiency characterized by recurrent sinopulmonary and herpes simplex virus infections and poor responses to immunization.
Sequence similaritiesBelongs to the peptidase C14A family. Contains 2 DED (death effector) domains.
DomainIsoform 9 contains a N-terminal extension that is required for interaction with the BCAP31 complex.
Post-translational modificationsGeneration of the subunits requires association with the death-inducing signaling complex (DISC), whereas additional processing is likely due to the autocatalytic activity of the activated protease. GZMB and CASP10 can be involved in these processing events. Phosphorylated upon DNA damage, probably by ATM or ATR.
Western blot - Active human Caspase-8 full length protein (ab54117)
Sample: Recombinant caspase 8 protein ab54117 at 100 ng. Primary antibody: Cleaved caspase 8 at 1:1000. Secondary antibody: Goat polyclonal to Rabbit IgG ? H&L at 1:10000. Development: ECL for exposure 10 min. Predicted band size: 18 kDa. Observed band size: 24 kDa.
Developed using the ECL technique
Performed under reducing conditions.
References for Recombinant human Caspase-8 protein (ab54117)
has not yet been referenced specifically in any publications.
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