Recombinant
RabMAb

Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493)

Abreviews (1)Q&A (5)Specific References (47)

Overview

  • Product name
    Anti-IRF3 (phospho S386) antibody [EPR2346]
    See all IRF3 primary antibodies
  • Description
    Rabbit monoclonal [EPR2346] to IRF3 (phospho S386)
  • Host species
    Rabbit
  • Specificity
    Detects IRF3 when phosphorylated on Serine 386.
  • Tested applications
    Suitable for: WB, Dot blotmore details
    Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Phospho-specific peptide corresponding to residues surrounding Serine 386 of Human IRF3

  • Positive control
    • MCF-7 cells treated with Calyculin A.
  • General notes

    A trial size is available to purchase for this antibody.

    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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab76493 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 1/1000 - 1/10000. Predicted molecular weight: 47 kDa.
Dot blot 1/1000.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF,IHC-P or IP.

    • 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 - Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493)
        Western blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493)
        All lanes : Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493) at 1/20000 dilution

        Lane 1 : MCF7 cell lysate - untreated
        Lane 2 : MCF7 cell lysate – treated with Calyculin A

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

        Predicted band size: 47 kDa



        Blocking buffer - 5% NFDM/TBST

        Diluting buffer - 1% BSA

      • Dot Blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493)
        Dot Blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493)

        Dot blot analysis of IRF3 single phospho peptide pS386 (lane 1) and IRF3 non-phospho peptide (lane 2) with ab76493 at 1/1000. Blocking and diluting buffer was 5% NFDM/TBST. The secondary antibody used was ab97051 Peroxidase conjugated Goat Anti-Rabbit IgG, (H+L) at 1/100,000.

      References

      This product has been referenced in:
      • Oh C  et al. A central role for PI3K-AKT signaling pathway in linking SAMHD1-deficiency to the type I interferon signature. Sci Rep 8:84 (2018). Read more (PubMed: 29311560) »
      • Stabell AC  et al. Dengue viruses cleave STING in humans but not in nonhuman primates, their presumed natural reservoir. Elife 7:N/A (2018). Read more (PubMed: 29557779) »

      See all 47 Publications for this product

      Publishing research using ab76493? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

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      6 reviews or Q&A

      10
      Votes: Highest First

      Western blot abreview for Anti-IRF3 (phospho S386) antibody [EPR2346]

       Good
      Abreviews
      Application
      Western blot
      Loading amount
      25 µg
      Gel Running Conditions
      Reduced Denaturing
      Sample
      Mouse Cell lysate - whole cell (liver)
      Specification
      liver
      Treatment
      20 ug/ml poly(I:C)
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
      Read More
      abreview image
      Username

      Abcam user community

      Verified customer

      Submitted Dec 26 2014

      Hi, can you recommend an isotype control to Ab below that would be suitable for flow cytometry?

      https://www.abcam.com/irf3-phospho-s386-antibody-epr2346-ab76493.html

      Thank you for contacting us. The rabbit monoclonal isotyoe control ab128142 should be suitable for your purposes in Flow Cytometry as well. Do you also need any secondary antibodies for your experiment? We offer many fluorophore conjugated secondaries ...

      Read More

      Zum oberen Bild:
      Geblockt wurde mit 5 % BSA in PBST für 1h bei RT. Anschließend wurde der P-IRF3 AK von Abcam (1:500 in 5 % BSA/PBST) zugegeben und über Nacht inkubiert. Das Ergebnis zeigt den P-IRF3 Antikörper. Es wurde kein anderer Antikörper zu...

      Read More

      Vielen Dank für Ihre Antwort.

      Ich möchte Ihnen einen kostenlosen Ersatz oder eine Gutschrift anbieten, da der Antikörper nicht so funktioniert wie wir diesen auf unserem Datenblatt beschreiben. Dazu benötige ich allerdings das ungefähre Best...

      Read More

      Hallo,
      attatched you will finde some excamples of the western blots.
      1) Abcam product code ab
      Ab-76493
      2) Abcam order reference number or product batch number
      lot: GR15710-3
      3) Description of the problem
      No P-IRF3 de...

      Read More

      Vielen Dank für das Ausfüllen unseres Fragebogens.

      Ich habe nur noch zwei Fragen bezüglich Ihres Protokolls:


      1.Haben Sie einen Blot nur mit BSA geblockt und anschliessend den prim. Antikörper in TBST+ BSA inkubiert?
      Read More

      Inquiry: Dear Sir or Madam, I have bought the Phospho-IRF3 antibody. I´m working with human hepatocytes and in my experimental setup I included positive controls (LPS or Poly I:C treated human hepatocytes). It also didn´t work with human st...

      Read More

      Thank you for contacting us.

      I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly. We have not re...

      Read More

      Can you please explain why the ICC image shows positive staining in untreated MCF7 cells? The WB image shows negative results in these untreated cells.

      Thank you for your call yesterday and for your patience while I have been in touch with the lab regarding ab76493. The phospho-specificity of the antibody was determined by Western blot and dot blot. The fluorescent detection may be more sensitive than...

      Read More

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