Validated using a knockout cell line
Recombinant
RabMAb

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

Overview

  • Product name
    Anti-Pro Caspase-8 antibody [EPR162]
    See all Pro Caspase-8 primary antibodies
  • Description
    Rabbit monoclonal [EPR162] to Pro Caspase-8
  • Host species
    Rabbit
  • Specificity

    Theoretically, ab108333 should recognize p55/54, p43/41 and p18. However, in our internal tests, it only recognized pro caspase 8. We tested it side by side with ab32397. Both antibodies are KO validated, and ab32397 recognizes pro-caspase 8 and cleavage 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 within Human Pro Caspase-8 aa 350-450 (internal sequence). The exact sequence is proprietary.
    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

     

     

    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 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

    Images

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Pro 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 Pro 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-Pro 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
      All lanes : 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 Pro Caspase-8 in paraffin-embedded human tonsil tissue by Immunohistochemistry.

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Pro 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-Pro 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-Pro 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 Pro Caspase-8 in paraffin-embedded human hepatocellular carcinoma by Immunohistochemistry.

    References

    This product has been referenced in:
    • Liao Y  et al. Endoplasmic Reticulum Stress Induces Macrophages to Produce IL-1ß During Mycobacterium bovis Infection via a Positive Feedback Loop Between Mitochondrial Damage and Inflammasome Activation. Front Immunol 10:268 (2019). Read more (PubMed: 30846986) »
    • Pittala S  et al. Targeting Liver Cancer and Associated Pathologies in Mice with a Mitochondrial VDAC1-Based Peptide. Neoplasia 20:594-609 (2018). WB ; Mouse . Read more (PubMed: 29747160) »
    See all 5 Publications for this product

    Customer reviews and Q&As

    Filter by Application

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    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton-X100 in PBS

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Aug 09 2012

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