Validated using a knockout cell line
Recombinant
RabMAb

Anti-DDX58 antibody [EPR18629] (ab180675)

Overview

  • Product name
    Anti-DDX58 antibody [EPR18629]
    See all DDX58 primary antibodies
  • Description
    Rabbit monoclonal [EPR18629] to DDX58
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human DDX58 aa 250-450. The exact sequence is proprietary.
    Database link: O95786

  • Positive control
    • WB: 293, HeLa and Jurkat whole cell lysates; Human fetal kidney and stomach lysates. IP: Jurkat whole cell lysate.
  • General notes

     

     

    Sold under exclusive licence from Children's Medical Research Institute and Newcastle Innovation Ltd. Pyrimidyn is a trademark of Children's Medical Research Institute and Newcastle Innovation Ltd.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab180675 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 1/1000. Detects a band of approximately 107 kDa (predicted molecular weight: 107 kDa).
IP 1/100.

Target

  • Function
    Involved in innate immune defense against viruses. Upon interaction with intracellular dsRNA produced during viral replication, triggers a transduction cascade involving MAVS/IPS1, which results in the activation of NF-kappa-B, IRF3 and IRF7 and the induction of the expression of antiviral cytokines such as IFN-beta and RANTES (CCL5). Detects dsRNA produced from non-self dsDNA by RNA polymerase III, such as Epstein-Barr virus-encoded RNAs (EBERs). Essential for the production of interferons in response to RNA viruses including paramyxoviruses, influenza viruses, Japanese encephalitis virus and HCV.
  • Tissue specificity
    Present in vascular smooth cells (at protein level).
  • Sequence similarities
    Belongs to the helicase family.
    Contains 2 CARD domains.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
  • Domain
    The repressor domain controls homomultimerization and interaction with MAVS.
    The helicase domain is responsible for dsRNA recognition.
    The 2 CARD domains are responsible for interaction with and signaling through MAVS.
    The second CARD domain is the primary site for 'Lys-63'-linked ubiquitination.
  • Post-translational
    modifications
    Isgylated. Conjugated to ubiquitin-like protein ISG15 upon IFN-beta stimulation.
    Ubiquitinated. Undergoes 'Lys-63'-linked ubiquitination. Lys-172 is the critical site for TRIM25-mediated ubiquitination, for MAVS binding and to induce anti-viral signal transduction. Lys-154, Lys-164 and Lys-172 are critical sites for RNF135-mediated ubiquitination. Deubiquitinated by CYLD, a protease that selectively cleaves 'Lys-63'-linked ubiquitin chains.
  • Cellular localization
    Cytoplasm. Colocalized with TRIM25 at cytoplasmic perinuclear bodies.
  • Information by UniProt
  • Database links
  • Alternative names
    • Ddx58 antibody
    • DDX58_HUMAN antibody
    • DEAD (Asp Glu Ala Asp) box polypeptide 58 antibody
    • DEAD (Asp Glu Ala Asp/His) box polypeptide antibody
    • DEAD box protein 58 antibody
    • DEAD/H (Asp Glu Ala Asp/His) box polypeptide RIG1 antibody
    • DKFZp434J1111 antibody
    • DKFZp686N19181 antibody
    • FLJ13599 antibody
    • Probable ATP dependent RNA helicase DDX58 antibody
    • Probable ATP-dependent RNA helicase DDX58 antibody
    • Retinoic acid inducible gene 1 protein antibody
    • Retinoic acid-inducible gene 1 protein antibody
    • Retinoic acid-inducible gene I protein antibody
    • RIG I antibody
    • Rig-1 antibody
    • RIG-I antibody
    • RIG1 antibody
    • rigi antibody
    • RLR 1 antibody
    • RNA helicase antibody
    • RNA helicase RIG I antibody
    • SGMRT2 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: DDX58 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: K562 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab180675 observed at 107 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab180675 was shown to specifically react with DDX58 when DDX58 knockout samples were used. Wild-type and DDX58 knockout samples were subjected to SDS-PAGE. ab180675 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-DDX58 antibody [EPR18629] (ab180675) at 1/1000 dilution

    Lane 1 : 293 (Human epithelial cells from embryonic kidney) whole cell lysate
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 107 kDa
    Observed band size: 107 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-DDX58 antibody [EPR18629] (ab180675) at 1/1000 dilution + Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 107 kDa
    Observed band size: 107 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-DDX58 antibody [EPR18629] (ab180675) at 1/1000 dilution + Human fetal kidney lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 107 kDa
    Observed band size: 107 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-DDX58 antibody [EPR18629] (ab180675) at 1/1000 dilution + Human stomach lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 107 kDa
    Observed band size: 107 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • DDX58 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab180675 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab180675 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.

    Lane 1: Jurkat whole cell lysate 10ug (Input). Lane 2: ab180675 IP in Jurkat whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab180675 in Jurkat whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 30 seconds.

References

ab180675 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Western blot
Sample
Pig Cell lysate - whole cell (Porcine kidney cells (PK-15))
Gel Running Conditions
Reduced Denaturing (4-20)
Loading amount
10 µg
Treatment
Porcine IFN-alpha for 20hrs
Specification
Porcine kidney cells (PK-15)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Mr. Sam Hardy

Verified customer

Submitted Aug 08 2017

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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