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  1. Link

    rig-iddx58-antibody-epr18629-ab180675.pdf

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Epigenetics and Nuclear Signaling DNA / RNA RNA Processing Other
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (1) Submit a question References (6)

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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

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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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18629
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • Other
    • Epigenetics and Nuclear Signaling
    • Chromatin Binding Proteins
    • DNA / RNA binding
    • Immunology
    • Immune System Diseases
    • Antiviral Signaling

Associated products

  • Alternative Versions

    • Anti-RIG-I/DDX58 antibody [EPR18629] - BSA and Azide free (ab240230)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

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

    • Human DDX58 (RIG-I) knockout A549 cell line (ab267116)
    • Human DDX58 (RIG-I) knockout A549 cell line (ab267117)
  • KO cell lysates

    • Human DDX58 (RIG-I) knockout A549 cell lysate (ab257916)
    • Human DDX58 (RIG-I) knockout A549 cell lysate (ab257917)
  • Recombinant Protein

    • Recombinant Human RIG-I/DDX58 protein (ab132502)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

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.
  • Target information above from: UniProt accession O95786 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 23586 Human
    • Omim: 609631 Human
    • SwissProt: O95786 Human
    • Unigene: 190622 Human
    • 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)

    Protocols

    • Western blot protocols
    • Immunoprecipitation protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    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

    Show All Reviews Q&A
    Submit a review Submit a question

    Western blot abreview for Anti-DDX58 antibody [EPR18629]

    Good
    Abreviews
    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

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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