Validated using a knockout cell line
Recombinant
RabMAb

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

Overview

  • Product name

    Anti-IRF3 antibody [EPR2418Y] (HRP)
    See all IRF3 primary antibodies
  • Description

    Rabbit monoclonal [EPR2418Y] to IRF3 (HRP)
  • 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

    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 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

  • All lanes : Anti-IRF3 antibody [EPR2418Y] (HRP) (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.

  • 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

References

ab205443 has not yet been referenced specifically in any publications.

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