Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-DDX5 antibody [EPR7239] (ab126730)

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Western blot - Anti-DDX5 antibody [EPR7239] (ab126730)
  • Flow Cytometry - Anti-DDX5 antibody [EPR7239] (ab126730)
  • Western blot - Anti-DDX5 antibody [EPR7239] (ab126730)
  • Immunocytochemistry/ Immunofluorescence - Anti-DDX5 antibody [EPR7239] (ab126730)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX5 antibody [EPR7239] (ab126730)
  • Immunoprecipitation - Anti-DDX5 antibody [EPR7239] (ab126730)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX5 antibody [EPR7239] (ab126730)
  • Immunocytochemistry/ Immunofluorescence - Anti-DDX5 antibody [EPR7239] (ab126730)
  • Anti-DDX5 antibody [EPR7239] (ab126730)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR7239] to DDX5
  • Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-DDX5 antibody [EPR7239]
    See all DDX5 primary antibodies

  • Description

    Rabbit monoclonal [EPR7239] to DDX5

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat

  • Immunogen

    Synthetic peptide, corresponding to residues in Human DDX5 (UniProt P17844).

  • Positive control

    • HeLa, SH-SY5Y, A431, and HT29 cell lysates; Human bladder carcinoma and Human kidney tissues; HeLa cells. IP: Mouse brain 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.

    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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR7239

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab126730 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
1/10000 - 1/50000. Detects a band of approximately 70 kDa (predicted molecular weight: 69 kDa).
IP
1/10 - 1/100.
IHC-P (2)
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
Use at an assay dependent concentration.
ICC/IF
1/100 - 1/250.
Notes
WB
1/10000 - 1/50000. Detects a band of approximately 70 kDa (predicted molecular weight: 69 kDa).
IP
1/10 - 1/100.
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
Use at an assay dependent concentration.
ICC/IF
1/100 - 1/250.

Target

  • Function

    RNA-dependent ATPase activity. The rate of ATP hydrolysis is highly stimulated by single-stranded RNA. May be involved in pre-mRNA splicing.

  • Sequence similarities

    Belongs to the DEAD box helicase family. DDX5/DBP2 subfamily.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.

  • Post-translational
    modifications

    Arg-502 is dimethylated, probably to asymmetric dimethylarginine.

  • Cellular localization

    Nucleus > nucleolus.
  • Information by UniProt

  • Database links

    • Alternative names

      • ATP dependent RNA helicase DDX5 antibody
      • DDX 5 antibody
      • Ddx5 antibody
      • DDX5_HUMAN antibody
      • DEAD (Asp Glu Ala Asp) box helicase 5 antibody
      • DEAD (Asp Glu Ala Asp) box polypeptide 5 antibody
      • DEAD box 5 antibody
      • DEAD box protein 5 antibody
      • DEAD/H (Asp Glu Ala Asp/His) box polypeptide 5 (RNA helicase, 68kD) antibody
      • G17P1 antibody
      • HELR antibody
      • HLR1 antibody
      • HUMP68 antibody
      • P68 antibody
      • p68 RNA helicase antibody
      • Probable ATP dependent RNA helicase DDX5 antibody
      • Probable ATP-dependent RNA helicase DDX5 antibody
      • RNA helicase p68 antibody
      see all

    Images

    • Western blot - Anti-DDX5 antibody [EPR7239] (ab126730)
      Western blot - Anti-DDX5 antibody [EPR7239] (ab126730)

      Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: DDX5 knockout HAP1 whole cell lysate (20 µg)
      Lane 3: MCF7 whole cell lysate (20 µg)
      Lane 4: HeLa whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab126730 observed at 69 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab126730 was shown to specifically react with DDX5 in wild-type cells as signal was lost in DDX5 knockout cells. Wild-type and DDX5 knockout samples were subjected to SDS-PAGE. Ab126730 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 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 1/20000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry - Anti-DDX5 antibody [EPR7239] (ab126730)
      Flow Cytometry - Anti-DDX5 antibody [EPR7239] (ab126730)

      ab126730 staining DDX5 in the human cell line HeLa (Human cervix adenocarcinoma epithelial cell) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/200. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

      Isoytype control: Rabbit monoclonal IgG (Black) Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue).

    • Western blot - Anti-DDX5 antibody [EPR7239] (ab126730)
      Western blot - Anti-DDX5 antibody [EPR7239] (ab126730)
      All lanes : Anti-DDX5 antibody [EPR7239] (ab126730) at 1/10000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : SH-SY5Y cell lysate
      Lane 3 : A431 cell lysate
      Lane 4 : HT29 cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 69 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-DDX5 antibody [EPR7239] (ab126730)
      Immunocytochemistry/ Immunofluorescence - Anti-DDX5 antibody [EPR7239] (ab126730)

      Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y (human neuroblastoma) cells labelling DDX5 (green) with purified ab126730 at 1/250. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as a nuclear counterstain.  

      Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX5 antibody [EPR7239] (ab126730)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX5 antibody [EPR7239] (ab126730)

      ab126730, at a dilution of 1/250, staining DDX5 in paraffin-embedded Human bladder carcinoma tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-DDX5 antibody [EPR7239] (ab126730)
      Immunoprecipitation - Anti-DDX5 antibody [EPR7239] (ab126730)

      Purified ab126730 at 1/20 dilution (1µg) immunoprecipitating DDX5 in Mouse brain lysate.
      Lane 1 (input): Mouse brain lysate 10µg
      Lane 2 (+): ab126730 + Mouse brain lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab126730 in Mouse brain lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/20,000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 69 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX5 antibody [EPR7239] (ab126730)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX5 antibody [EPR7239] (ab126730)

      ab126730, at a dilution of 1/250, staining DDX5 in paraffin-embedded Human kidney tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-DDX5 antibody [EPR7239] (ab126730)
      Immunocytochemistry/ Immunofluorescence - Anti-DDX5 antibody [EPR7239] (ab126730)
      ab126730, at a dilution of 1/100, staining DDX5 in HeLa cells by Immunofluorescence.
    • Anti-DDX5 antibody [EPR7239] (ab126730)
      Anti-DDX5 antibody [EPR7239] (ab126730)

    References (9)

    Publishing research using ab126730? Please let us know so that we can cite the reference in this datasheet.

    ab126730 has been referenced in 9 publications.

    • Spadotto V  et al. PRMT1-mediated methylation of the microprocessor-associated proteins regulates microRNA biogenesis. Nucleic Acids Res 48:96-115 (2020). PubMed: 31777917
    • Wang T  et al. LINC01116 promotes tumor proliferation and neutrophil recruitment via DDX5-mediated regulation of IL-1ß in glioma cell. Cell Death Dis 11:302 (2020). PubMed: 32358484
    • Wu J  et al. DDX5-targeting fully human monoclonal autoantibody inhibits proliferation and promotes differentiation of acute promyelocytic leukemia cells by increasing ROS production. Cell Death Dis 11:552 (2020). PubMed: 32690860
    • Marino MM  et al. Interactome mapping defines BRG1, a component of the SWI/SNF chromatin remodeling complex, as a new partner of the transcriptional regulator CTCF. J Biol Chem 294:861-873 (2019). PubMed: 30459231
    • Hoffmann N  et al. DGCR8 Promotes Neural Progenitor Expansion and Represses Neurogenesis in the Mouse Embryonic Neocortex. Front Neurosci 12:281 (2018). PubMed: 29760646
    • Dai L  et al. High expression of ALDOA and DDX5 are associated with poor prognosis in human colorectal cancer. Cancer Manag Res 10:1799-1806 (2018). PubMed: 29988738
    • Weise SC  et al. FOXG1 Regulates PRKAR2B Transcriptionally and Posttranscriptionally via miR200 in the Adult Hippocampus. Mol Neurobiol N/A:N/A (2018). PubMed: 30539330
    • Booy EP  et al. Comprehensive analysis of the BC200 ribonucleoprotein reveals a reciprocal regulatory function with CSDE1/UNR. Nucleic Acids Res 46:11575-11591 (2018). PubMed: 30247708
    • Choudhury NR  et al. RNA-binding activity of TRIM25 is mediated by its PRY/SPRY domain and is required for ubiquitination. BMC Biol 15:105 (2017). WB . PubMed: 29117863

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DDX5 antibody [EPR7239]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Small intestine)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH9
    Permeabilization
    No
    Specification
    Small intestine
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 23°C
    Fixative
    Formaldehyde
    Read More

    Mr. Victor Bernal

    Verified customer

    Submitted Nov 03 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-DDX5 antibody [EPR7239]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Tonsil)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH9
    Permeabilization
    No
    Specification
    Tonsil
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 2.5% · Temperature: 23°C
    Fixative
    Formaldehyde
    Read More

    Mr. Victor Bernal

    Verified customer

    Submitted Nov 03 2020

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