Human Caspase-8 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human cleaved Caspase-8 in ELISA.
  • Source
    Monoclonal Mouse IgG1 Clone # 746109
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    KLH-coupled Caspase-8 synthetic peptide
    CGIPVETD
    Accession # Q14790
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Immunocytochemistry
    8-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunocytochemistry
Caspase‑8 in Jurkat Human Cell Line. Caspase‑8 was detected in immersion fixed Jurkat human acute T cell leukemia cell line treated for 4 hours with staurosporin using Mouse Anti-Human Caspase‑8 Polyclonal Antibody (Catalog # MAB8135) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
Background: Caspase-8

Caspase-8 (Cysteine-aspartic acid protease 8/Casp8a; also named MCH5, FLICA and MACH alpha 1) is a 28 kDa member of the peptidase C14A family of enzymes (1, 2, 3). It is widely expressed and is considered an initiating caspase for the apoptotic cascade (4). Caspase-8 acts on a wide variety of substrates, including procaspases-3, 4, 6, 7, 9 and 10, c-FLIPL and procaspase‑8 itself (1, 5, 6). Human procaspase-8a is a 54‑56 kDa, 479 amino acid (aa) protein (4, 7, 8, 9). It contains two N-terminal death domains (aa 1‑177), followed by a catalytic site that utilizes His317Gly318 plus Cys360. Normally, it is an inactive, cytosolic monomer (1, 10, 11). But following death-domain (DD) containing receptor oligomerization, Caspase-8 is recruited to the death-inducing signaling complex (DISC) that forms around the death domains of the oligomerized receptor (12). FADD/CAP-1 is recruited first, followed by procaspase-8/CAP-4 and, possibly, c-FLIPL and procaspase‑10 (12). The recruitment, or concentration, of procaspase-8 induces homodimerization. This act alone is sufficient for activation. However, the activity level is modest at best, and appears to be directed towards either itself, or c-FLIPL, which is known to form a functional heterodimer with procaspase-8 (5, 11). When directed towards itself, autocleavage occurs first between Asp374Ser375, generating a 43 kDa (p43) N-terminal (aa 1‑374) and an 11 kDa C‑terminal (aa 375‑479) fragment. The C‑terminus is further cleaved between Asp384Leu385 to generate a mature p10 subunit (aa 385‑479). The p43 subunit is next cleaved twice, once between Asp216Ser217, and again between Asp210Ser211 to generate a 26 kDa DD-containing prodomain (aa 1‑210) with an additional 18 kDa mature p18 subunit (aa 217‑374) (12). p18 and p10 noncovalently associate to form a 28 kDa heterodimer, which subsequently associates with another p18:p10 heterodimer to form an active, mature Caspase-8 molecule. This leaves the DISC to act on downstream apoptotic procaspases. In the event procaspase-8 comes to the DISC complexed with c‑FLIPL, c‑FLIPL will be cleaved by procaspase-8, generating a p43 fragment that is analogous to the Caspase-8 p43 subunit. This fragment, however, appears not to be an intermediate in a proteolytic cascade. Rather, it serves as a functional subunit, interacting with TRAF2 and activating NF kappa B. This may account for many of the nonapoptotic activities associated with Caspase-8 (5, 6, 13). Mature human and mouse Caspase-8a heterodimers are 73% aa identical (14).

  • References:
    1. Chowdhury, I. et al. (2008) Comp. Biochem. Physiol. B 151:10.
    2. Boatright, K.M. & G.S. Salvesen (2003) Curr. Opin. Cell Biol. 15:725.
    3. Launay, S. et al. (2005) Oncogene 24:5137.
    4. Srinivasula, S.M. et al. (1996) Proc. Natl. Acad. Sci. USA 93:14486.
    5. Hughes, M.A. et al. (2009) Mol. Cell 35:265.
    6. Lamkanfi, M. et al. (2007) Cell Death Differ. 14:44.
    7. Fernandes-Alnemri, T. et al. (1996) Proc. Natl. Acad. Sci. USA 93:7464.
    8. Boldin, M.P. et al. (1996) Cell 85:803.
    9. Muzio, M. et al. (1996) Cell 85:817.
    10. Donepudi, M. et al. (2003) Mol. Cell 11:543.
    11. Boatright, K.M. et al. (2003) Mol. Cell 11:529.
    12. Golks, A. et al. (2006) Cell Death Differ. 13:489.
    13. Scaffidi, C. et al. (1997) J. Biol. Chem. 272:26953.
    14. Sakamaki, K. et al. (1998) Eur. J. Biochem. 253:399.
  • Entrez Gene IDs:
    841 (Human); 12370 (Mouse)
  • Alternate Names:
    AIS; androgen receptor; CASP8; Caspase8; Caspase-8; DHTRTFM; Dihydrotestosterone receptorHYSP1; HUMARA; Mch5; NR3C4KD; Nuclear receptor subfamily 3 group C member 4; SMAX1SBMA; spinal and bulbar muscular atrophy
Related Research Areas
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Mouse HRP-DAB Cell & Tissue Staining Kit

CTS002 14
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....
Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 Isotype Control

Ctrl MAB002 138  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Mouse IgG1 Isotype Control

Ctrl MAB002R 1
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....
Secondary Antibodies
Description Application Cat# Citations Images  

Mouse IgG HRP-conjugated Antibody

WB HAF007 26  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 10  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Mouse IgG Antibody

WB AF007 4
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Have you used Human Caspase-8 Antibody?

Submit a review and receive a $25US/€18/£15/$25CAN amazon gift card if you include an image - $10US/€7/£6/$10CAN Amazon card for reviews without an image. Limited to verified customers in USA, Canada and Europe.

Order Details

Contact Information

Calculators

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

=
÷
R&D Systems Guarantee

Monoclonal
Antibody
Panels For
Screening

Learn More
Monoclonal Antibodies

Associated Pathways

Feedback

Customer Information