Human/Mouse Caspase-8 Antibody

(4 citations)   
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human and mouse Caspase-8 in Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Caspase-8
    Ser234-Asp496
    Accession # AAC50645
  • 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
  • Western Blot
    0.5 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
Detection of Human and Mouse Caspase‑8 by Western Blot. Western blot shows lysates of Jurkat human leukemic T cell line and DA3 mouse myeloma cell line treated with 1 µM staurosporine for the indicated time. PVDF membrane was probed with 0.5 µg/mL Goat Anti-Human/Mouse Caspase‑8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF705) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands for Caspase‑8 precursor were detected at approximately 60 kDa (as indicated in upper panal) and specific bands for cleaved Caspase-8 were detected at approximately 14 - 18 kDa (as indicated in lower panal). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 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
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Multiplexed, targeted profiling of single-cell proteomes and transcriptomes in a single reaction
    Genome Biol, 2016;17(1):188.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC Fluorescence
  2. Ectodomain shedding of EGFR ligands and TNFR1 dictates hepatocyte apoptosis during fulminant hepatitis in mice.
    Authors: Murthy A, Defamie V, Smookler DS
    J. Clin. Invest., 2010;120(8):2731-44.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  3. Ellipticine derivative NSC 338258 represents a potential new antineoplastic agent for the treatment of multiple myeloma.
    Authors: Tian E, Landowski TH, Stephens OW, Yaccoby S, Barlogie B, Shaughnessy JD
    Mol. Cancer Ther., 2008;7(3):500-9.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. The B30.2 domain of pyrin, the familial Mediterranean fever protein, interacts directly with caspase-1 to modulate IL-1beta production.
    Authors: Chae JJ, Wood G, Masters SL, Richard K, Park G, Smith BJ, Kastner DL
    Proc. Natl. Acad. Sci. U.S.A., 2006;103(26):9982-7.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
Isotype Controls
Description Application Cat# Citations Images  

Normal Goat IgG Control

Ctrl AB-108-C 191  
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Secondary Antibodies
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Goat IgG HRP-conjugated Antibody

WB, Simple Western HAF109 19  
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Goat IgG HRP-conjugated Antibody

WB HAF017 15  
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Goat IgG (H+L) PE-conjugated Antibody

Flow F0107 4  
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Donkey Anti-Goat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL001 9
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Donkey Anti-Goat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL003 5
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Goat IgG (H+L) APC-conjugated Antibody

Flow F0108 6  
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Goat IgG Horseradish Peroxidase-conjugated Antibody

WB HAF019 6  
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Donkey Anti-Goat IgG Antibody

WB AF109 6
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Donkey Anti-Goat IgG Biotinylated Antibody

WB BAF109 3
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Goat IgG VisUCyte HRP Polymer

IHC VC004  
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Goat IgG (H+L) Fluorescein-conjugated Antibody

Flow F0109 2  
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Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL002
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Rabbit Anti-Goat IgG (H+L) Affinity Purified PAb, X Absorbed

WB, ELISA R-401-C-ABS
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Chicken Anti-Goat IgG Biotinylated Antibody

WB BAF019
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Rabbit Anti-Goat IgG Biotinylated Antibody

WB BAF017
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Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

Flow F0124
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