Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Caspase-8 antibody [E7] (ab32397)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E7] to Caspase-8
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Caspase-8 antibody [E7]
    See all Caspase-8 primary antibodies

  • Description

    Rabbit monoclonal [E7] to Caspase-8

  • Host species

    Rabbit

  • Specificity

    The antibody should recognize both pro-form (55kDa) and p18 cleaved-form of Caspase-8.

  • Tested Applications & Species

    Application Species
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: Wild-type/Wild-type treated with Staurosporin HAP1, Jurkat, SH-SY5Y, IM-9, and HeLa (ab150035) whole cell lysates.

  • General notes

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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

  • Western blot - Anti-Caspase-8 antibody [E7] (ab32397)
    Western blot - Anti-Caspase-8 antibody [E7] (ab32397)
    All lanes : Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : CASP8 knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : SH-SY5Y cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

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



    Lanes 1- 4: Merged signal (red and green). Green - ab32397 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab32397 was shown to react with Caspase-8 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264958 (knockout cell lysate ab256857) was used. Wild-type HeLa and CASP8 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32397 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Caspase-8 antibody [E7] (ab32397)
    Western blot - Anti-Caspase-8 antibody [E7] (ab32397)
    All lanes : Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : Wild-type HAP1 whole cell lysate treated with Staurosporin
    Lane 3 : Caspase-8 knockout HAP1 whole cell lysate
    Lane 4 : Caspase-8 knockout HAP1 whole cell lysate treated with Staurosporin

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 55 kDa



    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 1/500 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-Caspase-8 antibody [E7] (ab32397)
    Western blot - Anti-Caspase-8 antibody [E7] (ab32397)
    All lanes : Anti-Caspase-8 antibody [E7] (ab32397) at 1/1000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : IM-9 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution

    Predicted band size: 55 kDa



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-Caspase-8 antibody [E7] (ab32397)
    Western blot - Anti-Caspase-8 antibody [E7] (ab32397)
    Anti-Caspase-8 antibody [E7] (ab32397) at 1/500 dilution (unpurified) + HeLa cell lysate at 10 µg

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

  • Anti-Caspase-8 antibody [E7] (ab32397)
    Anti-Caspase-8 antibody [E7] (ab32397)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Alternative products to Anti-Caspase-8 antibody [E7] (ab32397)

© 2021 Astana Biomed Group LLP. All rights reserved.