Validated using a knockout cell line
Recombinant
RabMAb

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

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

    Suitable for: IP, WBmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human APAF1 aa 1-350. The exact sequence is proprietary.
    Database link: O14727

  • 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

    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 ab234436 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
IP 1/30.
WB 1/1000. Predicted molecular weight: 141 kDa.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IHC-P.
  • Target

    • Function

      Oligomeric Apaf-1 mediates the cytochrome c-dependent autocatalytic activation of pro-caspase-9 (Apaf-3), leading to the activation of caspase-3 and apoptosis. This activation requires ATP. Isoform 6 is less effective in inducing apoptosis.
    • Tissue specificity

      Ubiquitous. Highest levels of expression in adult spleen and peripheral blood leukocytes, and in fetal brain, kidney and lung. Isoform 1 is expressed in heart, kidney and liver.
    • Sequence similarities

      Contains 1 CARD domain.
      Contains 1 NB-ARC domain.
      Contains 13 WD repeats.
    • Domain

      The CARD domain mediates interaction with APIP.
    • Cellular localization

      Cytoplasm.
    • Information by UniProt
    • Database links

    • Alternative names

      • APAF 1 antibody
      • Apaf-1 antibody
      • APAF_HUMAN antibody
      • Apaf1 antibody
      • Apoptotic peptidase activating factor 1 antibody
      • Apoptotic protease activating factor 1 antibody
      • Apoptotic protease activating factor antibody
      • Apoptotic protease-activating factor 1 antibody
      • CED 4 antibody
      • CED4 antibody
      • KIAA0413 antibody
      see all

    Images

    • 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.

    • 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.

    • 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.

    • 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.

    References

    ab234436 has not yet been referenced specifically in any publications.

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