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    products/primary-antibodies/apaf1-antibody-epr21112-102-ab234436.pdf

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Cell Biology Apoptosis Mitochondrial
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • SDS
  • Certificate of Compliance
Submit a review Submit a question References (4)

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

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Protein
Product image
Recombinant Human APAF1 protein (Tagged) (ab198064)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

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. 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR21112-102
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Mitochondrial
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Cytochrome C
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Cytochrome C
    • Cancer
    • Cell Death
    • Apoptosis
    • Mitochondrial
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (ab254248)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human APAF1 protein (Tagged) (ab198064)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

Applications

The Abpromise guarantee

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.
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.
  • Target information above from: UniProt accession O14727 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 317 Human
    • Entrez Gene: 11783 Mouse
    • Entrez Gene: 78963 Rat
    • Omim: 602233 Human
    • SwissProt: O14727 Human
    • SwissProt: O88879 Mouse
    • SwissProt: Q9EPV5 Rat
    • Unigene: 728891 Human
    • Unigene: 220289 Mouse
    • Unigene: 64522 Rat
    see all
  • 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

  • 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)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (4)

Publishing research using ab234436? Please let us know so that we can cite the reference in this datasheet.

ab234436 has been referenced in 4 publications.

  • Zhang W  et al. l-Borneol ameliorates cerebral ischaemia by downregulating the mitochondrial calcium uniporter-induced apoptosis cascade in pMCAO rats. J Pharm Pharmacol 73:272-280 (2021). PubMed: 33793797
  • Sheng LX  et al. Design, Synthesis, and Evaluation of Novel 2-Methoxyestradiol Derivatives as Apoptotic Inducers Through an Intrinsic Apoptosis Pathway. Biomolecules 10:N/A (2020). PubMed: 31936880
  • Lin Y  et al. Psoralen alleviates high glucose-induced HK-2 cell injury by inhibition of Smad 2 signaling via upregulation of microRNA 874. BMC Pharmacol Toxicol 21:52 (2020). PubMed: 32698907
  • Gao Y  et al. Downregulation of CDC20 Increases Radiosensitivity through Mcl-1/p-Chk1-Mediated DNA Damage and Apoptosis in Tumor Cells. Int J Mol Sci 21:N/A (2020). PubMed: 32932732

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