Overview

  • Product name

    Anti-IRF3 antibody [EP2419Y] - BSA and Azide free
    See all IRF3 primary antibodies
  • Description

    Rabbit monoclonal [EP2419Y] to IRF3 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cytmore details
    Unsuitable for: ICC
  • Species reactivity

    Reacts with: Human
  • Immunogen

    A synthetic peptide corresponding to residues on the C terminus of human IRF3.

  • Positive control

    • U937, HeLa, MCF7 and Jurkat cell lysates; human tonsil tissue.
  • General notes

    ab201810 is the carrier-free version of ab76409 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Ab201810 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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

Our Abpromise guarantee covers the use of ab201810 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
IP
IHC-P
Flow Cyt
  • Application notes
    Flow Cyt: 1/10.
    IHC-P: 1/100 - 1/250. Antigen retrieval is recommended before commencing with IHC staining protocol.
    IP: 1/20.
    WB: 1/1000 - 1/2000. Predicted molecular weight: 47 kDa.

    Is unsuitable for ICC.

    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • 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

    • This WB data was generated using the same anti-IRF3 antibody clone, EP2419Y, in a different buffer formulation (cat# ab76409).

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: IRF3 knockout HAP1 cell lysate (20 µg) 
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: Jurkat cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab76409 observed at 50 kDa. Red - loading control, ab8245, observed at 37kDa.


      ab76409 was shown to react with IRF3 in wild-type HAP1 cells alond with additional cross-reactive bands. No band was observed when IRF3 knockout samples were used. Wild-type and IRF3 knockout samples were subjected to SDS-PAGE. ab76409 and ab8245 (loading control to GAPDH) were both 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 hour at room temperature before imaging.

    • Flow cytometric analysis of permeabilized U937 cells using anti- IRF-3 RabMAb (red) (ab76409) or a rabbit IgG (negative) (green)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76409).

    • This IHC data was generated using the same anti-IRF3 antibody clone, EP2419Y, in a different buffer formulation (cat# ab76409).

      Immunohistochemical analysis of IRF3 in paraffin embedded human tonsils using ab76409 at a 1/100 dilution.

      Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

      Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

    References

    ab201810 has not yet been referenced specifically in any publications.

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