Recombinant Anti-Caspase-8 antibody [E6] (ab32125)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E6] to Caspase-8
- Suitable for: WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Caspase-8 antibody [E6]
See all Caspase-8 primary antibodies -
Description
Rabbit monoclonal [E6] to Caspase-8 -
Host species
Rabbit -
Specificity
ab32125 should recognize all splice isoforms of Caspase-8. -
Tested applications
Suitable for: WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Caspase-8 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: Q14790 -
Epitope
ab32125 reacts with an epitope located in the N terminal region of caspase-8. -
Positive control
- WB: HeLa, SH-SY5Y, Jurkat (ab7899) and HAP1 whole cell lysates.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E6 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32125 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 |
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WB | (3) |
1/3000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
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Notes |
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WB
1/3000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa). |
Target
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Function
Most 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 specificity
Isoform 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 disease
Defects 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 similarities
Belongs to the peptidase C14A family.
Contains 2 DED (death effector) domains. -
Domain
Isoform 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. -
Cellular localization
Cytoplasm. - 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
- Apoptosis related cysteine peptidase antibody
see all
Images
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All lanes : Anti-Caspase-8 antibody [E6] (ab32125) at 1/3000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CASP8 knockout HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 55 kDaLanes 1- 4: Merged signal (red and green). Green - ab32125 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32125 was shown to react with Caspase-8 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264958 (knockout cell lysate ab256857) was used. Wild-type HeLa and CASP8 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32125 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 3000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Caspase-8 antibody [E6] (ab32125)
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : Caspase-8 knockout HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 55 kDaLanes 1 and 2: Merged signal (red and green). Green - ab32125 observed at 55 kDa. Red - loading control, ab8226, observed at 42 kDa.
ab32125 was shown to recognize Caspase-8 when Caspase-8 knockout samples were used, along with additional cross-reactive bands. Wild-type and Caspase-8 knockout samples were subjected to SDS-PAGE. ab32125 and ab8226 (loading control to beta actin) were diluted 1/3000 and 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
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Anti-Caspase-8 antibody [E6] (ab32125) at 1/3000 dilution + Jurkat cell lysate
Predicted band size: 55 kDa
Observed band size: 55 kDa
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (26)
ab32125 has been referenced in 26 publications.
- Holmgren C et al. Induction of Breast Cancer Cell Apoptosis by TRAIL and Smac Mimetics: Involvement of RIP1 and cFLIP. Curr Issues Mol Biol 44:4803-4821 (2022). PubMed: 36286042
- Humphreys LM et al. A revised model of TRAIL-R2 DISC assembly explains how FLIP(L) can inhibit or promote apoptosis. EMBO Rep 21:e49254 (2020). PubMed: 32009295
- Stöhr D et al. Stress-induced TRAILR2 expression overcomes TRAIL resistance in cancer cell spheroids. Cell Death Differ 27:3037-3052 (2020). PubMed: 32433558
- Dittmann J et al. Next-generation hypomethylating agent SGI-110 primes acute myeloid leukemia cells to IAP antagonist by activating extrinsic and intrinsic apoptosis pathways. Cell Death Differ 27:1878-1895 (2020). PubMed: 31831875
- Wu T et al. Expression of receptor interacting protein 3 and mixed lineage kinase domain-like protein-key proteins in necroptosis is upregulated in ulcerative colitis. Ann Palliat Med 8:483-489 (2019). PubMed: 31431023