Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

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 - BSA and Azide free
  • Suitable for: IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free
    See all APAF1 primary antibodies

  • Description

    Rabbit monoclonal [EPR21112-102] to APAF1 - BSA and Azide free

  • 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

    • IP: ab234436 IP in HEK-293T whole cell lysate. WB: Wild-type HAP1 whole cell lysate. HEK-293T, HeLa and A20 whole cell lysate.

  • General notes

    Ab254248 is the carrier-free version of ab234436. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab254248 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

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

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab234436).

  • Western blot - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (ab254248)
    Western blot - Anti-APAF1 antibody [EPR21112-102] - BSA and Azide free (ab254248)
    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab234436).

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

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