Recombinant Anti-Caspase-8 antibody [E7] (ab32397)
![Western blot - Anti-Caspase-8 antibody [E7] (ab32397)](https://www.abcam.com/ps/products/32/ab32397/Images/ab32397-397506-rabbit-monoclonal-epr2418y-to-irf3-western-blot-hela-jurkat-sh-sy5y-lysates.jpg "Western blot - Anti-Caspase-8 antibody [E7] (ab32397)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E7] 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 [E7]
See all Caspase-8 primary antibodies -
Description
Rabbit monoclonal [E7] to Caspase-8 -
Host species
Rabbit -
Specificity
The antibody should recognize both pro-form (55kDa) and p18 cleaved-form of Caspase-8. -
Tested applications
Suitable for: WBmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Caspase-8 aa 200-300 (N terminal). The exact sequence is proprietary.
Database link: Q14790 -
Positive control
- WB: Wild-type/Wild-type treated with Staurosporin HAP1, Jurkat, SH-SY5Y, IM-9, and HeLa (ab150035) 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.
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.
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: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E7 -
Isotype
IgG -
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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KO cell lines
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KO cell lysates
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KO cell pellets
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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 | (2) |
1/500 - 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
For unpurified use at 1/500. |
| Notes |
|---|
|
WB
1/500 - 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa). For unpurified use at 1/500. |
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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Western blot - Anti-Caspase-8 antibody [E7] (ab32397)All lanes : Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 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 - ab32397 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32397 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. ab32397 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 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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![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)All lanes : Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Wild-type HAP1 whole cell lysate treated with Staurosporin
Lane 3 : Caspase-8 knockout HAP1 whole cell lysate
Lane 4 : Caspase-8 knockout HAP1 whole cell lysate treated with Staurosporin
Lysates/proteins at 20 µg per lane.
Predicted band size: 55 kDaLanes 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 1/500 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) + HeLa cell lysate at 10 µg
Predicted band size: 55 kDa
Observed band size: 55 kDa -
![Anti-Caspase-8 antibody [E7] (ab32397)](https://www.abcam.com/ps/products/32/ab32397/Images/ab32397-6-benefits-of-recombinant-antibodies.png)
Anti-Caspase-8 antibody [E7] (ab32397)
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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Datasheet download
References (23)
ab32397 has been referenced in 23 publications.
- Zhang Y et al. Long Non-coding RNA CASC15 Promotes Intrahepatic Cholangiocarcinoma Possibly through Inducing PRDX2/PI3K/AKT Axis. Cancer Res Treat 53:184-198 (2021). PubMed: 33017884
- Zhao A et al. lncRNA TUSC7 inhibits osteosarcoma progression through the miR-181a/RASSF6 axis. Int J Mol Med 47:583-594 (2021). PubMed: 33416181
- Albahde MAH et al. CBL0137 administration suppresses human hepatocellular carcinoma cells proliferation and induces apoptosis associated with multiple cell death related proteins. Neoplasma 67:547-556 (2020). PubMed: 32202904
- Ou R et al. HPV16 E6 oncoprotein-induced upregulation of lncRNA GABPB1-AS1 facilitates cervical cancer progression by regulating miR-519e-5p/Notch2 axis. FASEB J 34:13211-13223 (2020). PubMed: 32844486
- Yao J et al. Rapamycin inhibits proliferation and apoptosis of retinoblastoma cells through PI3K/AKT signaling pathway. Oncol Lett 19:2950-2956 (2020). PubMed: 32218850
