Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-APAF1 antibody [EPR21112-102] (ab234436)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21112-102] to APAF1
  • Suitable for: IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-APAF1 antibody [EPR21112-102]
    See all APAF1 primary antibodies

  • Description

    Rabbit monoclonal [EPR21112-102] to APAF1

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IP
    Human
    WB
    Rat
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal heart amd fetal kidney tissue lysate. Rat spleen and liver tissue lysate. Wild-type HAP1 whole cell lysate. HEK-293T, HeLa and A20 whole cell lysate. IP: ab234436 IP in HEK-293T whole cell lysate.

  • General notes

    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.

Properties

Images

  • Western blot - Anti-APAF1 antibody [EPR21112-102] (ab234436)
    Western blot - Anti-APAF1 antibody [EPR21112-102] (ab234436)
    All lanes : Anti-APAF1 antibody [EPR21112-102] (ab234436) at 1/1000 dilution

    Lane 1 : Human fetal heart tissue lysate
    Lane 2 : Human fetal kidney tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

    Predicted band size: 141 kDa


    Exposure time: 3 minutes


    Blocking/dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-APAF1 antibody [EPR21112-102] (ab234436)
    Western blot - Anti-APAF1 antibody [EPR21112-102] (ab234436)
    All lanes : Anti-APAF1 antibody [EPR21112-102] (ab234436) at 1/1000 dilution

    Lane 1 : Rat spleen tissue lysate
    Lane 2 : Rat liver tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 141 kDa



    Lanes 2 was developed using a higher sensitivity ECL substrate.

    Exposure times: Lane 1: 169 secs; Lane 2: 3 mins.

    Blocking/dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-APAF1 antibody [EPR21112-102] (ab234436)
    Western blot - Anti-APAF1 antibody [EPR21112-102] (ab234436)
    All lanes : Anti-APAF1 antibody [EPR21112-102] (ab234436) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : APAF1 knockout HAP1 whole cell lysate
    Lane 3 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen), whole cell lysate
    Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 5 : A20 (mouse reticulum sarcoma B lymphocyte), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 141 kDa



    Ab234436 was shown to specifically react with APAF1 in wild-type HAP1 cells as signal was lost in APAF1 knockout cells. Wild-type and APAF1 knockout samples were subjected to SDS-PAGE. ab234436 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging.

    Lanes 1,2 and 5 were developed using a higher sensitivity ECL substrate.

    Exposure times: Lanes 1-2: 136 secs; Lanes 3-4: 3 mins; Lane 5: 136 secs.

    Blocking/dilution buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-APAF1 antibody [EPR21112-102] (ab234436)
    Immunoprecipitation - Anti-APAF1 antibody [EPR21112-102] (ab234436)

    APAF1 was immunoprecipitated from 0.35 mg HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate with ab234436 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab234436 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.

    Lane 1: HEK-293T whole cell lysate 10 µg (Input).
    Lane 2: ab234436 IP in HEK-293T whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab234436 in HEK-293T whole cell lysate.

    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 10 seconds.

  • Anti-APAF1 antibody [EPR21112-102] (ab234436)
    Anti-APAF1 antibody [EPR21112-102] (ab234436)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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