Validated using a knockout cell lineRecombinantRabMAb

Recombinant HRP Anti-IRF3 antibody [EPR2418Y] (ab205443)

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Western blot - HRP Anti-IRF3 antibody [EPR2418Y] (ab205443)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-IRF3 antibody [EPR2418Y] (ab205443)
  • HRP Anti-IRF3 antibody [EPR2418Y] (ab205443)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • HRP Rabbit monoclonal [EPR2418Y] to IRF3
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Human
  • Conjugation: HRP

Overview

  • Product name

    HRP Anti-IRF3 antibody [EPR2418Y]
    See all IRF3 primary antibodies

  • Description

    HRP Rabbit monoclonal [EPR2418Y] to IRF3

  • Host species

    Rabbit

  • Conjugation

    HRP

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse

  • Immunogen

    Synthetic peptide within Human IRF3 aa 50-150. The exact sequence is proprietary.
    Database link: Q14653

  • Positive control

    • WB: HeLa, Jurkat and wild-type HAP1 whole cell lysates. IHC-P: Normal human tonsil tissue.

  • 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab205443 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
IHC-P 1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 51 kDa (predicted molecular weight: 47 kDa).

Target

  • Function

    Mediates interferon-stimulated response element (ISRE) promoter activation. Functions as a molecular switch for antiviral activity. DsRNA generated during the course of an viral infection leads to IRF3 phosphorylation on the C-terminal serine/threonine cluster. This induces a conformational change, leading to its dimerization, nuclear localization and association with CREB binding protein (CREBBP) to form dsRNA-activated factor 1 (DRAF1), a complex which activates the transcription of genes under the control of ISRE. The complex binds to the IE and PRDIII regions on the IFN-alpha and IFN-beta promoters respectively. IRF-3 does not have any transcription activation domains.

  • Tissue specificity

    Expressed constitutively in a variety of tissues.

  • Sequence similarities

    Belongs to the IRF family.
    Contains 1 IRF tryptophan pentad repeat DNA-binding domain.

  • Post-translational
    modifications

    Constitutively phosphorylated on many serines residues. C-terminal serine/threonine cluster is phosphorylated in response of induction by IKBKE and TBK1. Ser-385 and Ser-386 may be specifically phosphorylated in response to induction. An alternate model propose that the five serine/threonine residues between 396 and 405 are phosphorylated in response to a viral infection. Phosphorylation, and subsequent activation of IRF3 is inhibited by vaccinia virus protein E3.
    Ubiquitinated; ubiquitination involves RBCK1 leading to proteasomal degradation. Polyubiquitinated; ubiquitination involves TRIM21 leading to proteasomal degradation.
    ISGylated by HERC5 resulting in sustained IRF3 activation and in the inhibition of IRF3 ubiquitination by disrupting PIN1 binding. The phosphorylation state of IRF3 does not alter ISGylation.

  • Cellular localization

    Cytoplasm. Nucleus. Shuttles between cytoplasmic and nuclear compartments, with export being the prevailing effect. When activated, IRF3 interaction with CREBBP prevents its export to the cytoplasm.
  • Information by UniProt

  • Database links

    • Alternative names

      • IIAE7 antibody
      • Interferon regulatory factor 3 antibody
      • IRF 3 antibody
      • IRF-3 antibody
      • IRF3 antibody
      • IRF3_HUMAN antibody
      • MGC94729 antibody
      see all

    Images

    • Western blot - HRP Anti-IRF3 antibody [EPR2418Y] (ab205443)
      Western blot - HRP Anti-IRF3 antibody [EPR2418Y] (ab205443)
      All lanes : HRP Anti-IRF3 antibody [EPR2418Y] (ab205443) at 1/2000 dilution

      Lane 1 : HeLa whole cell lysate (ab150035) at 10 µg
      Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg
      Lane 3 : Wild-type HAP1 cell lysate at 20 µg
      Lane 4 : IRF3 knockout HAP1 cell lysate at 20 µg

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 47 kDa
      Observed band size: 51 kDa
      why is the actual band size different from the predicted?


      Exposure time: 20 minutes


      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab205443 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-IRF3 antibody [EPR2418Y] (ab205443)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-IRF3 antibody [EPR2418Y] (ab205443)

      IHC image of IRF3 staining in a section of formalin-fixed paraffin-embedded normal human tonsil tissue*, performed on a Leica BONDTM. The section was pre-treated using heat mediated antigen retrieval with TRIS/EDTA (pH9, epitope retrieval solution 2) for 20mins.  The section was then incubated with ab205443, 1/50 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • HRP Anti-IRF3 antibody [EPR2418Y] (ab205443)
      HRP Anti-IRF3 antibody [EPR2418Y] (ab205443)

    References (0)

    Publishing research using ab205443? Please let us know so that we can cite the reference in this datasheet.

    ab205443 has not yet been referenced specifically in any publications.

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