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  1. Link

    irf3-antibody-epr2418y-ab68481.pdf

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Immunology Innate Immunity Cytokines Interferons
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IRF3 antibody [EPR2418Y] (ab68481)

  • Datasheet
  • SDS
Reviews (2)Q&A (2)References (41)

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Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Immunocytochemistry/ Immunofluorescence - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Flow Cytometry (Intracellular) - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Flow Cytometry (Intracellular) - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EPR2418Y] (ab68481)
  • Anti-IRF3 antibody [EPR2418Y] (ab68481)

Key features and details

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

Conjugates logo Related conjugates and formulations

Carrier Free HRP PE

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Anti-IRF3 (phospho S386) antibody [EPR2346] - BSA and Azide free (ab182859)
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Human IRF3 knockout HeLa cell line (ab255345)
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Human IRF3 knockout A549 cell line (ab267097)

View more associated products

Overview

  • Product name

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

    Rabbit monoclonal [EPR2418Y] to IRF3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human IRF3 aa 50-150. The exact sequence is proprietary.
    Database link: Q14653
    (Peptide available as ab203561)

  • Positive control

    • WB: A549, HeLa, Jurkat, THP-1, Daudi, HepG2 whole cell lysates. Human fetal heart and kidney lysates. Mouse heart and spleen lysates, NIH/3T3 whole cell lysates. IHC-P: Human tonsil, Human squamous cell carcinoma of cervix, Mouse spleen. ICC/IF: HeLa cells Flow: U937 cells, HAP1-wt cells.
  • 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.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR2418Y
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interferons
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Immunology
    • Innate Immunity
    • TLR Signaling

Associated products

  • Alternative Versions

    • Anti-IRF3 antibody [EPR2418Y] - BSA and Azide free (ab201809)
    • HRP Anti-IRF3 antibody [EPR2418Y] (ab205443)
    • PE Anti-IRF3 antibody [EPR2418Y] (ab209919)
  • Assay kits

    • IRF3 Transcription Factor Assay Kit (Colorimetric) (ab207210)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Immunizing Peptide (Blocking)

    • IRF3 peptide (ab203561)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human IRF3 knockout HeLa cell line (ab255345)
    • Human IRF3 knockout A549 cell line (ab267097)
    • Human IRF3 knockout A549 cell line (ab267098)
  • KO cell lysates

    • Human IRF3 knockout A549 cell lysate (ab256953)
    • Human IRF3 knockout A549 cell lysate (ab256954)
    • Human IRF3 knockout HeLa cell lysate (ab263784)
  • Recombinant Protein

    • Recombinant Human IRF3 protein (ab205216)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab68481 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
Flow Cyt (Intra)
1/160.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  MeOH fixationis recommended.

ICC/IF
1/100.
WB (1)
1/1000. Detects a band of approximately 51 kDa (predicted molecular weight: 47 kDa).Can be blocked with IRF3 peptide (ab203561).
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
Flow Cyt (Intra)
1/160.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  MeOH fixationis recommended.

ICC/IF
1/100.
WB
1/1000. Detects a band of approximately 51 kDa (predicted molecular weight: 47 kDa).Can be blocked with IRF3 peptide (ab203561).
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Application notes
Is unsuitable for 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.
  • Target information above from: UniProt accession Q14653 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3661 Human
    • Entrez Gene: 54131 Mouse
    • Omim: 603734 Human
    • SwissProt: Q14653 Human
    • SwissProt: P70671 Mouse
    • Unigene: 289052 Human
    • Unigene: 75254 Human
    • Unigene: 3960 Mouse
    • 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 antibody [EPR2418Y] (ab68481)
      Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      All lanes : Anti-IRF3 antibody [EPR2418Y] (ab68481) at 1/1000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : MCF7 cell lysate
      Lane 3 : Wild-type HeLa cell lysate
      Lane 4 : IRF3 knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 47 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab68481 observed at 50 kDa. Red - loading control, ab8245 observed at 37 kDa.  

       ab68481 was shown to react with IRF3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255345 (knockout cell lysate ab263784) was used. Wild-type and IRF3 knockout samples were subjected to SDS-PAGE. ab68481 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

       

    • Immunocytochemistry/ Immunofluorescence - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Immunocytochemistry/ Immunofluorescence - Anti-IRF3 antibody [EPR2418Y] (ab68481)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling IRF3 with ab68481 at 1/100 dilution. Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/400 dilution was used as the secondary antibody (green). The confocal image shows cytoplasmic on HeLa cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
      The negative controls are as follows;
      1. ab68481 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
      2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

       

    • Flow Cytometry (Intracellular) - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Flow Cytometry (Intracellular) - Anti-IRF3 antibody [EPR2418Y] (ab68481)

      Overlay histogram showing HAP1 wildtype (green line) and HAP1-IRF3 knockout cells (red line) stained with ab68481. The cells were fixed with 80% methanol (5 min) (left pannel) or 4% formaldehyde (10 min) (right pannel), and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab68481, 0.1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) presorbed (ab150081) at 1/2000 dilution for 30 min at 22°C. A rabbit IgG isotype control antibody (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-IRF3 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. 

      Note: We recommend fixing cells using MeOHinstead of PFA toget optimal results. 

       

       

    • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      All lanes : Anti-IRF3 antibody [EPR2418Y] (ab68481) at 1/1000 dilution

      Lane 1 : Wild-type A549 cell lysate
      Lane 2 : IRF3 knockout A549 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 47 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?



      Lanes 1 - 2: Merged signal (red and green). Green - ab68481 observed at 50 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

      ab68481 was shown to react with IRF3 in wild-type A549 cells in western blot with loss of signal observed in IRF3 knockout cell line ab267098 (IRF3 knockout cell lysate ab256954). Wild-type and IRF3 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab68481 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      All lanes : Anti-IRF3 antibody [EPR2418Y] (ab68481) at 1/1000 dilution

      Lane 1 : Wild-type A549 cell lysate
      Lane 2 : IRF3 knockout A549 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 47 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?



      Lanes 1 - 2: Merged signal (red and green). Green - ab68481 observed at 50 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

      ab68481 was shown to react with IRF3 in wild-type A549 cells in western blot with loss of signal observed in IRF3 knockout cell line ab267097 (IRF3 knockout cell lysate ab256953). Wild-type and IRF3 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab68481 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      All lanes : Anti-IRF3 antibody [EPR2418Y] (ab68481) at 1/1000 dilution

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

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 47 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab68481 observed at 50 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab68481 was shown to react with IRF3 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when IRF3 knockout samples were examined. Wild-type and IRF3 knockout samples were subjected to SDS-PAGE. ab68481 and ab8245 (loading control to GAPDH) were both diluted to 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.

    • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      All lanes : Anti-IRF3 antibody [EPR2418Y] (ab68481) at 1/1000 dilution

      Lane 1 : Human fetal heart lysate
      Lane 2 : Human fetal kidney lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 47 kDa



      Blocking and Dilution buffer: 5% NFDM/TBST

    • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      All lanes : Anti-IRF3 antibody [EPR2418Y] (ab68481) at 1/10000 dilution

      Lane 1 : THP-1 (Human monocytic leukemia cells) whole cell lysates
      Lane 2 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysates
      Lane 3 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysates

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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



      Blocking and Dilution buffer: 5% NFDM/TBST

    • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      All lanes : Anti-IRF3 antibody [EPR2418Y] (ab68481) at 1/10000 dilution

      Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
      Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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



      Blocking and Dilution buffer: 5% NFDM/TBST

    • Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Western blot - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      All lanes : Anti-IRF3 antibody [EPR2418Y] (ab68481) at 1/1000 dilution

      Lane 1 : Mouse heart lysate
      Lane 2 : Mouse spleen lysate
      Lane 3 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 47 kDa
      Observed band size: 47 kDa



      Blocking and Dilution buffer: 5% NFDM/TBST

       

      The slightly smaller molecular mass observed in mouse than in human is supported by literature.

    • Flow Cytometry (Intracellular) - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Flow Cytometry (Intracellular) - Anti-IRF3 antibody [EPR2418Y] (ab68481)

      Intracellular Flow Cytometry analysis of 2% paraformaldehyde fixed U937 (Human histiocytic lymphoma cells)cells labeling IRF3 with ab68481 at 1/160 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control is rabbit monoclonal IgG (black line). The unlabeled control is cells without incubation with primary and secondary antibodies (blue line). 

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EPR2418Y] (ab68481)

      Immunohistochemical analysis of paraffin-embedded Human tonsil labeling IRF3 with ab68481at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EPR2418Y] (ab68481)

      Immunohistochemical analysis of paraffin-embedded Human squamous cell carcinoma of cervix labeling IRF3 with ab68481at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EPR2418Y] (ab68481)

      Immunohistochemical analysis of paraffin-embedded Mouse spleen labeling IRF3 with ab68481at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Anti-IRF3 antibody [EPR2418Y] (ab68481)
      Anti-IRF3 antibody [EPR2418Y] (ab68481)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (41)

    Publishing research using ab68481? Please let us know so that we can cite the reference in this datasheet.

    ab68481 has been referenced in 41 publications.

    • Banerjee A  et al. Experimental and natural evidence of SARS-CoV-2-infection-induced activation of type I interferon responses. iScience 24:102477 (2021). PubMed: 33937724
    • Moustaqil M  et al. SARS-CoV-2 proteases PLpro and 3CLpro cleave IRF3 and critical modulators of inflammatory pathways (NLRP12 and TAB1): implications for disease presentation across species. Emerg Microbes Infect 10:178-195 (2021). PubMed: 33372854
    • Kim JS  et al. Multi-Functional MPT Protein as a Therapeutic Agent against Mycobacterium tuberculosis. Biomedicines 9:N/A (2021). PubMed: 34068051
    • Zhang T  et al. microRNA-365 inhibits YAP through TLR4-mediated IRF3 phosphorylation and thereby alleviates gastric precancerous lesions. Cancer Cell Int 20:549 (2020). PubMed: 33292210
    • Liao Z  et al. Heat-killed Salmonella Typhimurium protects mice against carbon ion radiation. J Int Med Res 48:300060520924256 (2020). PubMed: 33021413
    View all Publications for this product

    Customer reviews and Q&As

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    Western blot abreview for Anti-IRF3 antibody [EPR2418Y]

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    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (human hepatocytes)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    15 µg
    Specification
    human hepatocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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    Abcam user community

    Verified customer

    Submitted Feb 01 2018

    Flow Cytometry abreview for Anti-IRF3 antibody [EPR2418Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (primary human macrophages and CD4+ T cells)
    Permeabilization
    Yes - BD Cytofix/cytoperm kit
    Gating Strategy
    all live cells
    Specification
    primary human macrophages and CD4+ T cells
    Preparation
    Cell harvesting/tissue preparation method: Monocyte-derived macrophages were prepared from human PBMCs CD4+ T cells were separated from human PBMCs
    Sample buffer: 0.5%BSA/PBS
    Fixation
    Paraformaldehyde
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    Radiana Trifonova

    Verified customer

    Submitted May 27 2016

    Question

    I'd just like to ask whether this antibody is known to be suitable for the detection of native mouse IRF3 for an IRF3 dimerization assay.

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    Abcam community

    Verified customer

    Asked on Feb 08 2013

    Answer

    ab68481 has been tested in human hela cells and MCF7 lysates and can be used as positive controls for your experiment to assess the efficacy of the antibody. Abcam provides a whole range of protocols to test both antibodies.

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    Abcam Scientific Support

    Answered on Feb 08 2013

    Question


    For our assay, we are attempting to detect IRF3 activation via dimerization in mouse macrophage nuclear and cytoplasmic lysates run on a native gel. It is important for us that the antibody is able to detect both the monomer and dimer forms at comparable efficiencies in order to make a comparison.

    In your experience, has either ab25950 or ab68481 been successfully used with mouse samples?

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    Abcam community

    Verified customer

    Asked on Feb 12 2013

    Answer

    ab25950 and ab68481, has been tested in mouse whole cell lysates however,we have only tested the antibody in a reduced, denatured Western blot.
    We are not sure whether it will bind to a dimer in a native Western blot. It is possible, but it's also possible that the epitope of the antibody (located in the C-terminus) will be obscured in the native or dimerized form, and in that case the antibody would not bind to the protein. We can not be sure of this until it is tested.

    Read More

    Abcam Scientific Support

    Answered on Feb 12 2013

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