Validated using a knockout cell line
Recombinant
RabMAb

Anti-Caspase-8 antibody [EPR162] (ab108333)

Overview

  • Product name
    Anti-Caspase-8 antibody [EPR162]
    See all Caspase-8 primary antibodies
  • Description
    Rabbit monoclonal [EPR162] to Caspase-8
  • Tested applications
    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Caspase-8 aa 200-300 (internal sequence).
    Database link: Q14790

  • Positive control
    • Jurkat, IM9, HepG2, HeLa, and HL60 cell lysates; Human tonsil tissue and Human hepatocellular carcinoma tissue; HeLa cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    Alternative versions available:

    Anti-Caspase-8 antibody (Alexa Fluor® 488) [EPR162] (ab206068)

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Properties

Applications

Our Abpromise guarantee covers the use of ab108333 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 1/1000 - 1/10000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
IHC-P 1/100 - 1/250. Antigen retrieval is recommended.
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • Target

    • 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
      modifications
      Generation 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
    • Database links
    • Alternative names
      • ALPS2B antibody
      • Amyotrophic lateral sclerosis 2 chromosomal region candidate gene 12 protein antibody
      • Apoptotic cysteine protease antibody
      • Apoptotic protease Mch-5 antibody
      • Apoptotic protease Mch5 antibody
      • CAP4 antibody
      • CASP-8 antibody
      • CASP8 antibody
      • CASP8_HUMAN antibody
      • Caspase 8 antibody
      • Caspase 8 apoptosis related cysteine peptidase antibody
      • Caspase-8 subunit p10 antibody
      • CED 3 antibody
      • FADD Like ICE antibody
      • FADD-homologous ICE/CED-3-like protease antibody
      • FADD-like ICE antibody
      • FLICE antibody
      • FLJ17672 antibody
      • ICE-like apoptotic protease 5 antibody
      • MACH alpha 1/2/3 protein antibody
      • MACH antibody
      • MACH beta 1/2/3/4 protein antibody
      • MCH5 antibody
      • MGC78473 antibody
      • MORT1 associated ced 3 homolog antibody
      • MORT1-associated CED-3 homolog antibody
      • OTTHUMP00000163717 antibody
      • OTTHUMP00000163720 antibody
      • OTTHUMP00000163724 antibody
      • OTTHUMP00000163725 antibody
      • OTTHUMP00000165062 antibody
      • OTTHUMP00000165063 antibody
      • OTTHUMP00000165064 antibody
      • OTTHUMP00000206552 antibody
      • OTTHUMP00000206582 antibody
      see all

    Anti-Caspase-8 antibody [EPR162] images



    • Predicted band size : 55 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Caspase-8 knockout HAP1 cell lysate (20 µg)
      Lanes 1 and 2: Merged signal (red and green). Green - ab108333 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab108333 was shown to specifically react with Caspase-8 when Caspase-8 knockout samples were used. Wild-type and Caspase-8 knockout samples were subjected to SDS-PAGE. ab108333 and ab8245 (loading control to GAPDH) were both diluted 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.

    • All lanes : Anti-Caspase-8 antibody [EPR162] (ab108333) at 1/1000 dilution

      Lane 1 : HepG2 whole cell lysate
      Lane 2 : Rat liver tissue lysate
      Lane 3 : Mouse liver tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size : 55 kDa
      Observed band size : 55 kDa

      Lanes 1 - 3: Merged signal (red and green). Green - ab108333 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa. 

      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab108333 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

    • ab108333, at 1/100 dilution, staining Caspase-8 in paraffin-embedded human tonsil tissue by Immunohistochemistry.



    • Predicted band size : 55 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Caspase-8 knockout HAP1 cell lysate (20 µg)
      Lanes 1 and 2: Merged signal (red and green).

      Green - target observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.

      This western blot image is a comparison between ab108333 and a competitor's top cited mouse monoclonal antibody.

    • All lanes : Anti-Caspase-8 antibody [EPR162] (ab108333) at 1/1000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : IM9 cell lysate
      Lane 3 : HepG2 cell lysate
      Lane 4 : HL60 cell lysate

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 55 kDa
    • All lanes : Anti-Caspase-8 antibody [EPR162] (ab108333) at 1/1000 dilution

      Lane 1 : Jurkat cell lysate, treated with etoposide
      Lane 2 : Jurkat cell lysate
      Lane 3 : HeLa cell lysate, treated staurosporine
      Lane 4 : HeLa cell lysate

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 55 kDa
    • ab108333, at 1/100 dilution, staining Caspase-8 in paraffin-embedded human hepatocellular carcinoma by Immunohistochemistry.

    References for Anti-Caspase-8 antibody [EPR162] (ab108333)

    This product has been referenced in:
    • Wang J  et al. Menin mediates Tat-induced neuronal apoptosis in brain frontal cortex of SIV-infected macaques and in Tat-treated cells. Oncotarget 8:18082-18094 (2017). WB ; Rhesus monkey . Read more (PubMed: 28178646) »
    • Stutz N  et al. The Fas apoptotic pathway in cutaneous T-cell lymphomas: Frequent expression of phenotypes associated with resistance to apoptosis. J Am Acad Dermatol : (2012). IHC ; Human . Read more (PubMed: 22884443) »

    See all 2 Publications for this product

    Product Wall


    The immunogen sequence is considered proprietary information. The sequence is a 30-amino acid sequence within the larger range of amino acids from 200 - 250 of the human protein sequence that is given in the UniProt entry at this link: http://www....

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton-X100 in PBS
    Username

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Aug 09 2012

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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