RecombinantRabMAb

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

Reviews (1)Q&A (5)References (87)
Western blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493)
  • Western blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493)
  • Dot Blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493)
  • Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2346] to IRF3 (phospho S386)
  • Suitable for: WB, Dot blot
  • Reacts with: Human

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

    IRF3 (phospho S386) antibody (ab76493) detects IRF3 when phosphorylated on Serine 386.

  • Tested applications

    Suitable for: WB, Dot blotmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide corresponding to Human IRF3 (phospho S386).
    Database link: Q14653

  • Positive control

    • WB: MCF7 cells treated with Calyculin A.

  • General notes

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

     

    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.

    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.

Properties

Applications

The Abpromise guarantee

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)
1/1000 - 1/10000. Predicted molecular weight: 47 kDa.
Dot blot
1/1000.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 47 kDa.
Dot blot
1/1000.

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 0.2 µg/ml (purified)

      Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) treated with calyculin A for 45 minutes whole cell lysates
      Lane 3 : MCF7 (Human breast adenocarcinoma epithelial cell) treated with calyculin A for 45 minutes whole cell lysates. Then the membrane was incubated with phosphatase

      Lysates/proteins at 15 µg per lane.

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

      Predicted band size: 47 kDa



      Blocking and diluting buffer: 5% NFDM/TBST

      We are unsure to define the extra band at 37KD.

    • 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 (Unpurified)

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

      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 unpurified 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.

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

    References (87)

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

    ab76493 has been referenced in 87 publications.

    • Zhong L  et al. Phosphorylation of cGAS by CDK1 impairs self-DNA sensing in mitosis. Cell Discov 6:26 (2020). PubMed: 32351706
    • Zhu Q  et al. A synthetic STING agonist inhibits the replication of human parainfluenza virus 3 and rhinovirus 16 through distinct mechanisms. Antiviral Res 183:104933 (2020). PubMed: 32949635
    • Tong W  et al. Resveratrol inhibits LPS-induced inflammation through suppressing the signaling cascades of TLR4-NF-?B/MAPKs/IRF3. Exp Ther Med 19:1824-1834 (2020). PubMed: 32104238
    • Guerra J  et al. Lysyl-tRNA synthetase produces diadenosine tetraphosphate to curb STING-dependent inflammation. Sci Adv 6:eaax3333 (2020). PubMed: 32494729
    • Zhang H  et al. ADAR1 Facilitates KSHV Lytic Reactivation by Modulating the RLR-Dependent Signaling Pathway. Cell Rep 31:107564 (2020). PubMed: 32348766
    View all Publications for this product

    Customer reviews and Q&As

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

     Good
    Abreviews
    abreview image
    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

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    Verified customer

    Submitted Dec 26 2014

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