Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Caspase-8 antibody [E7] - BSA and Azide free (ab232046)

Overview

  • Product name
    Anti-Caspase-8 antibody [E7] - BSA and Azide free
    See all Caspase-8 primary antibodies
  • Description
    Rabbit monoclonal [E7] to Caspase-8 - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IFmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Caspase-8 aa 200-300. The exact sequence is proprietary.
    Database link: Q14790

  • Positive control
    • WB: Wild type/ Wild type treated with Staurosporin HAP1 whole cell lysate.
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®). 

    ab232046 is a PBS-only buffer format of ab32397. Please refer to ab32397 for recommended dilutions, protocols, and image data.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab232046 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 Use at an assay dependent concentration. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).

 

ICC/IF Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for Flow Cyt.
  • 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

    Images

    • ab32394 staining Caspase-8 in the K562 cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/150). ab150078(1/500) an anti-rabbit Alexa Fluor®555 was used as the secondary antibody. Nuclei were counterstained with DAPI.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32397).

    • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: Wild type HAP1 whole cell lysate treated with Staurosporin (20 µg)
      Lane 3: Caspase-8 knockout HAP1 whole cell lysate (20 µg)
      Lane 4: Caspase-8 knockout HAP1 whole cell lysate treated with Staurosporin (20 µg)

      Lanes 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 500 dilution and 1/20,000 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 1/20,000 dilution for 1 hour at room temperature before imaging.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32397).

       

    References

    ab232046 has not yet been referenced specifically in any publications.

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