Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Caspase-8 aa 200-300. A synthetic peptide corresponding to N-terminal residues of human Caspase-8 subunit p18 was used as immunogen. Database link: Q14790
Hela cell lysate
This product is a recombinant rabbit monoclonal antibody.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated "PUR" on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/5000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
For unpurified use at 1/500.
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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.
ab32394 staining Caspase-8 in the K562 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/150). ab150078(1/500) an anti-rabbit Alexa Fluor®555 was used as the secondary antibody. Nuclei were counterstained with DAPI.
Western blot - Anti-Caspase-8 antibody [E7] (ab32397)
All lanes : Anti-Caspase-8 antibody [E7] (ab32397) at 1/1000 dilution
Lane 1 : Jurkat cell lysate Lane 2 : HeLa cell lysate Lane 3 : IM-9 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution
Predicted band size : 55 kDa
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-Caspase-8 [E7] antibody (ab32397)
Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution (unpurified) + 10ug HeLa cell lysate.
Predicted band size : 55 kDa Observed band size : 55 kDa
References for Anti-Caspase-8 antibody [E7] (ab32397)
This product has been referenced in:
Altonsy MO et al. Differential induction of apoptosis in human colonic carcinoma cells (Caco-2) by Atopobium, and commensal, probiotic and enteropathogenic bacteria: mediation by the mitochondrial pathway. Int J Food Microbiol137:190-203 (2010).
Read more (PubMed: 20036023) »
Kaneko M et al. Prolonged low-dose administration of the cyclooxygenase-2 inhibitor celecoxib enhances the antitumor activity of irinotecan against neuroblastoma xenografts. Cancer Sci100:2193-201 (2009).
Read more (PubMed: 19673886) »