Caspase-8 (Cysteine-aspartic acid protease 8/Casp8; also MCH5 and FLICA) is a 28 kDa member of the peptidase C14A family of enzymes. It is known as the initiating caspase for the apoptotic cascade. Caspase-8 acts on procaspases-3, 4, 6, 7, 9 and 10, in addition to c-FLIPL and procaspase-8 itself. Human procaspase-8 alpha 4 is a 58 kDa (predicted), 496 amino acid (aa) protein and contains two N-terminal death domains plus a catalytic site that utilizes His334Gly335 plus Cys377. Following death domain-containing receptor activation, proteolysis generates a 28 kDa heterodimer. This includes a p18 subunit (aa 234-391; or 217-374 of the 8a/standard form) noncovalently linked to a p10 subunit (aa 402-496; or 385-479 of the 8a/standard form). Association with another p18/p10 heterodimer generates active caspase-8. There are two common procaspase-8 isoform variants. One 53 kDa isoform (8b) shows a deletion of aa 103-134, while a second 55 kDa (8a) isoform shows a deletion of aa 103-134 with a 15 aa insertion after Lys183. The p18 and p10 subunits are 68% and 82% aa identical, human to mouse, respectively.
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
Western Blot
Cited:
Western Blot, Immunocytochemistry
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG2A Clone # 84131
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Product Specifications
Immunogen
E. coli-derived recombinant human Caspase-8
aa 234-391
aa 234-391
Specificity
Detects human Caspase-8 precursor in Western blots and a 42 kDa doublet generated during apoptosis.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG2A
Scientific Data Images for Human Caspase-8 Antibody (84131)
Detection of Human Caspase‑8 by Western Blot.
Western blot shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with Human FAS Antigen Affinity-purified Polyclonal Antibody (Catalog # AF126) for indicated times. PVDF membrane was probed with 1 µg/mL of Human Caspase-8 Monoclonal Antibody (Catalog # MAB704), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Specific bands were detected for Caspase-8 at approximately 55 and 42 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.Applications for Human Caspase-8 Antibody (84131)
Application
Recommended Usage
Western Blot
1 µg/mL
Sample: Anti Fas-treated Jurkat human acute T cell leukemia cell line
Sample: Anti Fas-treated Jurkat human acute T cell leukemia cell line
Reviewed Applications
Read 2 reviews rated 4.5 using MAB704 in the following applications:
Formulation, Preparation, and Storage
Purification
Protein A or G purified from ascites
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: Caspase-8
Additional Caspase-8 Products
Product Documents for Human Caspase-8 Antibody (84131)
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Caspase-8 Antibody (84131)
For research use only
Related Research Areas
Citations for Human Caspase-8 Antibody (84131)
Customer Reviews for Human Caspase-8 Antibody (84131) (2)
4.5 out of 5
2 Customer Ratings
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Application: Western BlotSample Tested: MBA-MB-123 human breast cancer cell line and A549 human lung carcinoma cell lineSpecies: HumanVerified Customer | Posted 10/17/2018
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Application: Western BlotSample Tested: See PMID 23536519Species: HumanVerified Customer | Posted 02/25/2015
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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