Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PDCD4 antibody [EPR3432] - BSA and Azide free (ab247499)

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 - BSA and Azide free
  • Suitable for: IHC-P, ICC/IF, WB
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-PDCD4 antibody [EPR3432] - BSA and Azide free
    See all PDCD4 primary antibodies

  • Description

    Rabbit monoclonal [EPR3432] to PDCD4 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC/IF, WBmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, HAP1 and HEK293 cell lysates. ICC/IF: HeLa cells.

  • General notes

    ab247499 is the carrier-free version of ab79405 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

    Ab247499 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this 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.

Properties

Images

  • Western blot - Anti-PDCD4 antibody [EPR3432] - BSA and Azide free (ab247499)
    Western blot - Anti-PDCD4 antibody [EPR3432] - BSA and Azide free (ab247499)
    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?



    This data was developed using the same antibody clone in a different buffer formulation (ab79405).

      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] - BSA and Azide free (ab247499)
    Immunocytochemistry/ Immunofluorescence - Anti-PDCD4 antibody [EPR3432] - BSA and Azide free (ab247499)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab79405).

  • Western blot - Anti-PDCD4 antibody [EPR3432] - BSA and Azide free (ab247499)
    Western blot - Anti-PDCD4 antibody [EPR3432] - BSA and Azide free (ab247499)
    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



    This data was developed using the same antibody clone in a different buffer formulation (ab79405).

    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.

  • Anti-PDCD4 antibody [EPR3432] - BSA and Azide free (ab247499)
    Anti-PDCD4 antibody [EPR3432] - BSA and Azide free (ab247499)

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