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    pdcd4-antibody-epr3432-ab79405.pdf

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Cell Biology Cell Cycle Cell Cycle Inhibitors Other
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PDCD4 antibody [EPR3432] (ab79405)

  • Datasheet
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Western blot - Anti-PDCD4 antibody [EPR3432] (ab79405)
  • Immunocytochemistry/ Immunofluorescence - Anti-PDCD4 antibody [EPR3432] (ab79405)
  • Western blot - Anti-PDCD4 antibody [EPR3432] (ab79405)
  • Western blot - Anti-PDCD4 antibody [EPR3432] (ab79405)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDCD4 antibody [EPR3432] (ab79405)
  • 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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 555 Carrier Free PE

You may also be interested in

Knockout
Product image
Human PDCD4 knockout HeLa cell line (ab261833)

View more associated products

Overview

  • Product name

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

    Rabbit monoclonal [EPR3432] to PDCD4
  • Host species

    Rabbit
  • Tested applications

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

    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.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3432
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Other
    • Cell Biology
    • Apoptosis
    • Nucleus
    • Other
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Other
    • Cancer
    • Cell Death
    • Apoptosis
    • Other
    • Cancer
    • Cell Death
    • Apoptosis
    • Nucleus
    • Other

Associated products

  • Alternative Versions

    • PE Anti-PDCD4 antibody [EPR3432] (ab209301)
    • Alexa Fluor® 555 Anti-PDCD4 antibody [EPR3432] (ab215673)
    • Anti-PDCD4 antibody [EPR3432] - BSA and Azide free (ab247499)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human PDCD4 knockout HeLa cell line (ab261833)
  • KO cell lysates

    • Human PDCD4 knockout HeLa cell lysate (ab257278)
  • KO cell pellets

    • Human PDCD4 knockout HeLa cell pellet (ab279002)
  • Positive Controls

    • Jurkat cytoplasmic extract lysate (ab14842)
  • Recombinant Protein

    • Recombinant Human PDCD4 protein (ab172822)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab79405 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
WB
1/5000 - 1/10000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
IHC-P
1/1000 - 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF
1/500.
Notes
WB
1/5000 - 1/10000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
IHC-P
1/1000 - 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF
1/500.
Application notes
Is unsuitable for IP.

Target

  • Function

    Inhibits translation initiation and cap-dependent translation. May excert its function by hindering the interaction between EIF4A1 and EIF4G. Inhibits the helicase activity of EIF4A. Modulates the activation of JUN kinase. Down-regulates the expression of MAP4K1, thus inhibiting events important in driving invasion, namely, MAPK85 activation and consequent JUN-dependent transcription. May play a role in apoptosis. Tumor suppressor. Inhibits tumor promoter-induced neoplastic transformation. Binds RNA.
  • Tissue specificity

    Up-regulated in proliferative cells. Highly expressed in epithelial cells of the mammary gland. Reduced expression in lung cancer and colon carcinoma.
  • Sequence similarities

    Belongs to the PDCD4 family.
    Contains 2 MI domains.
  • Domain

    Binds EIF4A1 via both MI domains.
  • Post-translational
    modifications

    Polyubiquitinated, leading to its proteasomal degradation. Rapidly degraded in response to mitogens. Phosphorylation of the phosphodegron promotes interaction with BTRC and proteasomal degradation.
  • Cellular localization

    Nucleus. Cytoplasm. Shuttles between the nucleus and cytoplasm. Predominantly nuclear under normal growth conditions, and when phosphorylated at Ser-457. Exported from the nucleus in the absence of serum.
  • Target information above from: UniProt accession Q53EL6 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 27250 Human
    • Entrez Gene: 18569 Mouse
    • Omim: 608610 Human
    • SwissProt: Q53EL6 Human
    • SwissProt: Q61823 Mouse
    • Unigene: 711490 Human
    • Unigene: 1605 Mouse
    • Unigene: 375091 Mouse
    • Alternative names

      • Death up-regulated gene protein antibody
      • Dug antibody
      • H731 antibody
      • Ma3 antibody
      • MGC33046 antibody
      • MGC33047 antibody
      • Neoplastic transformation inhibitor antibody
      • Neoplastic transformation inhibitor protein antibody
      • Nuclear antigen H731 antibody
      • Nuclear antigen H731 like antibody
      • Nuclear antigen H731 like protein antibody
      • Nuclear antigen H731-like antibody
      • PDCD 4 antibody
      • Pdcd4 antibody
      • PDCD4_HUMAN antibody
      • Programmed cell death 4 antibody
      • programmed cell death 4 (neoplastic transformation inhibitor) antibody
      • Programmed cell death protein 4 antibody
      • Protein 197/15a antibody
      • Protein MA-3 antibody
      • Tis antibody
      • Topoisomerase-inhibitor suppressed protein antibody
      see all

    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)

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (8)

    Publishing research using ab79405? Please let us know so that we can cite the reference in this datasheet.

    ab79405 has been referenced in 8 publications.

    • Hale BJ  et al. Inhibition of germinal vesicle breakdown using IBMX increases microRNA-21 in the porcine oocyte. Reprod Biol Endocrinol 18:39 (2020). PubMed: 32393269
    • Patutina OA  et al. Mesyl phosphoramidate backbone modified antisense oligonucleotides targeting miR-21 with enhanced in vivo therapeutic potency. Proc Natl Acad Sci U S A 117:32370-32379 (2020). PubMed: 33288723
    • Hua R  et al. Ssc-miR-21-5p regulates endometrial epithelial cell proliferation, apoptosis and migration via the PDCD4/AKT pathway. J Cell Sci 133:N/A (2020). PubMed: 33097608
    • Chen T  et al. Involvement of androgen receptor (AR)/microRNA-21 axis in hypoxia/reoxygenation-induced apoptosis of mouse renal tubular epithelial cells. Am J Transl Res 11:5611-5622 (2019). PubMed: 31632533
    • Zhang Y  et al. Expression and function of Pdcd4 in mouse endometrium during early pregnancy. Reproduction 155:393-402 (2018). PubMed: 29459402
    • Patutina OA  et al. miRNases: Novel peptide-oligonucleotide bioconjugates that silence miR-21 in lymphosarcoma cells. Biomaterials 122:163-178 (2017). PubMed: 28126663
    • Liang X  et al. MicroRNA-16 suppresses the activation of inflammatory macrophages in atherosclerosis by targeting PDCD4. Int J Mol Med 37:967-75 (2016). PubMed: 26936421
    • Wright EC  et al. MicroRNA-21 and PDCD4 expression during in vitro oocyte maturation in pigs. Reprod Biol Endocrinol 14:21 (2016). PubMed: 27084064

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