Overview

  • Product name
    Anti-Caspase-8 antibody [E7] (HRP)
    See all Caspase-8 primary antibodies
  • Description
    Rabbit monoclonal [E7] to Caspase-8 (HRP)
  • Host species
    Rabbit
  • Conjugation
    HRP
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    A synthetic peptide corresponding to N-terminal residues of human Caspase-8 subunit p18 was used as immunogen.

  • Positive control
    • WB: HeLa whole cell lysate.
  • General notes

    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 ab194145 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 1/5000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).

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

  • All lanes : Anti-Caspase-8 antibody [E7] (HRP) (ab194145) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : CASP8 knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 55 kDa


    Exposure time: 20 minutes


    ab194145 was shown to specifically react with Caspase-8 in wild-type HAP1 cells as signal was lost in CASP8 knockout cells. Wild-type and CASP8 knockout samples were subjected to SDS-PAGE. ab194145 was incubated overnight at 4oC at 1/5000 dilution. Blots were developed with ECL technique.

  • Anti-Caspase-8 antibody [E7] (HRP) (ab194145) at 1/5000 dilution + HeLa whole cell lysate (ab150035) at 10 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

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


    Exposure time: 20 minutes


    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 3% milk before being incubated with ab194145 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

References

ab194145 has not yet been referenced specifically in any publications.

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