Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PDCD4 antibody [EPR3432] (ab79405)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3432] to PDCD4
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-PDCD4 antibody [EPR3432]
    See all PDCD4 primary antibodies

  • Description

    Rabbit monoclonal [EPR3432] to PDCD4

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human PDCD4 aa 400-500 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa, HAP1, and Jurkat whole cell lysate. IHC-P: Human breast carcinoma tissue. ICC/IF: HeLa cells

  • General notes

    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-PDCD4 antibody [EPR3432] (ab79405)
    Western blot - Anti-PDCD4 antibody [EPR3432] (ab79405)
    All lanes : Anti-PDCD4 antibody [EPR3432] (ab79405) at 1/5000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : PDCD4 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 52 kDa
    Observed band size: 51 kDa
    why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab79405 observed at 51 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab79405 was shown to react with PDCD4 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261833 (knockout cell lysate ab257278) was used. Wild-type HeLa and PDCD4 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. ab79405 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-PDCD4 antibody [EPR3432] (ab79405)
    Immunocytochemistry/ Immunofluorescence - Anti-PDCD4 antibody [EPR3432] (ab79405)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling PDCD4 with purified ab79405 at 1/500 dilution (4 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL) was used as the secondary antibody only control.

  • Western blot - Anti-PDCD4 antibody [EPR3432] (ab79405)
    Western blot - Anti-PDCD4 antibody [EPR3432] (ab79405)
    All lanes : Anti-PDCD4 antibody [EPR3432] (ab79405) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : PDCD4 knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : HEK293 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 52 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab79405 observed at 52 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab79405 was shown to recognize PDCD4 in wild-type HAP1 cells as signal was lost at the expected MW in PDCD4 knockout HAP1 cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PDCD4 knockout samples were subjected to SDS-PAGE. ab79405 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 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 1/10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-PDCD4 antibody [EPR3432] (ab79405)
    Western blot - Anti-PDCD4 antibody [EPR3432] (ab79405)
    All lanes : Anti-PDCD4 antibody [EPR3432] (ab79405) at 1/10000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size: 52 kDa
    Observed band size: 52 kDa
    Additional bands at: 30 kDa. We are unsure as to the identity of these extra bands.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDCD4 antibody [EPR3432] (ab79405)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDCD4 antibody [EPR3432] (ab79405)

    ab79405 at 1/1000 dilution staining PDCD4 in human breast carcinoma by Immunohistochemistry using paraffin-embedded tissue.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Anti-PDCD4 antibody [EPR3432] (ab79405)
    Anti-PDCD4 antibody [EPR3432] (ab79405)

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