Recombinant Anti-Caspase-8 antibody [E7] (ab32397)
- Datasheet
- References (10)
- Protocols
Overview
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Product nameAnti-Caspase-8 antibody [E7]
See all Caspase-8 primary antibodies -
DescriptionRabbit monoclonal [E7] to Caspase-8
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Host speciesRabbit
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SpecificityThe antibody should recognize both pro-form (55kDa) and p18 cleaved-form of Caspase-8.
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Tested applicationsSuitable for: WBmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivityReacts with: Human
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Immunogen
Synthetic peptide within Human Caspase-8 aa 200-300 (N terminal). The exact sequence is proprietary.
Database link: Q14790 -
Positive control
- HeLa whole cell lysate (ab150035)
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General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
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.
This product is a recombinant rabbit monoclonal antibody.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberE7
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IsotypeIgG
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Research areas
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab32397 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | 1/1000 - 1/5000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa). For unpurified use at 1/500. |
Target
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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.
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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.
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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.
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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. -
Cellular localizationCytoplasm.
- Information by UniProt
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Database links
- Entrez Gene: 841 Human
- Omim: 601763 Human
- SwissProt: Q14790 Human
- Unigene: 599762 Human
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Alternative names
- ALPS2B antibody
- Amyotrophic lateral sclerosis 2 chromosomal region candidate gene 12 protein antibody
- Apoptotic cysteine protease antibody
see all
Images
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![Western blot - Anti-Caspase-8 antibody [E7] (ab32397)](https://www.abcam.com/ps/products/32/ab32397/Images/ab32397-282490-anti-caspase-8-antibody-e7-western-blot.jpg)
Western blot - Anti-Caspase-8 antibody [E7] (ab32397)Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: HAP1 + Staurosproin knockout HAP1 whole cell lysate (20 µg)
Lane 3: CASP8 whole cell lysate (20 µg)
Lane 4: CASP8 + Staurosporin whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab32397 observed at 55, 43/41 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab32397 was shown to specifically react with HAP1 + Staurosproin when HAP1 + Staurosproin knockout samples were used. Wild-type and HAP1 + Staurosproin knockout samples were subjected to SDS-PAGE. Ab32397 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-Caspase-8 antibody [E7] (ab32397)](https://www.abcam.com/ps/products/32/ab32397/Images/ab32397-240569-anti-caspase-8-antibody-e7-western-blot.jpg)
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
All lanes : Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution
Predicted band size: 55 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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![Western blot - Anti-Caspase-8 antibody [E7] (ab32397)](https://www.abcam.com/ps/products/32/ab32397/Images/ab32397-44701-anti-caspase-8-antibody-e7-western-blot.jpg)
Western blot - Anti-Caspase-8 antibody [E7] (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
Datasheets and documents
References
This product has been referenced in:
- Zhang C et al. Growth of tyrosine kinase inhibitor-resistant Philadelphia-positive acute lymphoblastic leukemia: Role of bone marrow stromal cells. Oncol Lett 13:2059-2070 (2017). WB . Read more (PubMed: 28454362) »
- Jin P et al. Essential role of microRNA-650 in the regulation of B-cell CLL/lymphoma 11B gene expression following transplantation: A novel mechanism behind the acute rejection of renal allografts. Int J Mol Med 40:1840-1850 (2017). Read more (PubMed: 29039465) »
