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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)

Reviews (1) Submit a question References (6)
Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
  • Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
  • Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
  • Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
  • Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
  • Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
  • Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
  • Immunoprecipitation - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
  • Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18629] to RIG-I/DDX58
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-RIG-I/DDX58 antibody [EPR18629]
    See all RIG-I/DDX58 primary antibodies

  • Description

    Rabbit monoclonal [EPR18629] to RIG-I/DDX58

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A549, 293, HeLa and Jurkat whole cell lysates; Human fetal kidney and stomach lysates. IP: Jurkat whole cell 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.

Properties

Applications

The Abpromise guarantee

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)
1/1000. Detects a band of approximately 107 kDa (predicted molecular weight: 107 kDa).
IP
1/100.
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

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

      Lane 1 : Wild-type A549 cell lysate
      Lane 2 : DDX58 knockout A549 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

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



      Lanes 1 - 2: Merged signal (red and green). Green - ab180675 observed at 107 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

      ab180675 was shown to react with DDX58 in A549 wild-type cells in western blot with loss of signal observed in DDX58 knockout cell line ab267117 (DDX58 knockout cell lysate ab257917). Wild-type and DDX58 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab180675 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

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

      Lane 1 : Wild-type A549 cell lysate
      Lane 2 : DDX58 knockout A549 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

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



      Lanes 1 - 2: Merged signal (red and green). Green - ab180675 observed at 107 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

      ab180675 was shown to react with DDX58 in wild-type A549 cells in western blot with loss of signal observed in DDX58 knockout cell line ab267116 (DDX58 knockout cell lysate ab257916). Wild-type and DDX58 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab180675 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
      Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: RIG-I/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 RIG-I/DDX58 when RIG-I/DDX58 knockout samples were used. Wild-type and RIG-I/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.

    • Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
      Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
      All lanes : Anti-RIG-I/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.

    • Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
      Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
      Anti-RIG-I/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.

    • Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
      Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
      Anti-RIG-I/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.

    • Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
      Western blot - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
      Anti-RIG-I/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.

    • Immunoprecipitation - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
      Immunoprecipitation - Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)

      RIG-I/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 Detection Reagent (HRP) (ab131366), was used for detection 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.

    • Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)
      Anti-RIG-I/DDX58 antibody [EPR18629] (ab180675)

    References (6)

    Publishing research using ab180675? Please let us know so that we can cite the reference in this datasheet.

    ab180675 has been referenced in 6 publications.

    • Lu M  et al. N6-methyladenosine modification enables viral RNA to escape recognition by RNA sensor RIG-I. Nat Microbiol 5:584-598 (2020). PubMed: 32015498
    • Zhang X  et al. Foot-and-Mouth Disease Virus 3B Protein Interacts with Pattern Recognition Receptor RIG-I to Block RIG-I-Mediated Immune Signaling and Inhibit Host Antiviral Response. J Immunol 205:2207-2221 (2020). PubMed: 32917788
    • Liu G  et al. Inhibition of Ongoing Influenza A Virus Replication Reveals Different Mechanisms of RIG-I Activation. J Virol 93:N/A (2019). PubMed: 30602605
    • Chen YG  et al. N6-Methyladenosine Modification Controls Circular RNA Immunity. Mol Cell N/A:N/A (2019). PubMed: 31474572
    • Yi L  et al. Interferon regulatory factor 3 mediates Poly(I:C)-induced innate immune response and apoptosis in non-small cell lung cancer. Int J Oncol 52:1623-1632 (2018). PubMed: 29512705
    • Zhao Y  et al. RIG-I like receptor sensing of host RNAs facilitates the cell-intrinsic immune response to KSHV infection. Nat Commun 9:4841 (2018). PubMed: 30451863

    Customer reviews and Q&As

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    Western blot abreview for Anti-DDX58 antibody [EPR18629]

     Good
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MR. Sam Hardy

    Verified customer

    Submitted Aug 08 2017

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