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Rabbit Recombinant Monoclonal IRF3 antibody. Suitable for IHC-P, IP, WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 25 publications.

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Images

Western blot - Anti-IRF3 antibody [EP2419Y] (AB76409), expandable thumbnail
  • Western blot - Anti-IRF3 antibody [EP2419Y] (AB76409), expandable thumbnail
  • Immunoprecipitation - Anti-IRF3 antibody [EP2419Y] (AB76409), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EP2419Y] (AB76409), expandable thumbnail
  • Western blot - Anti-IRF3 antibody [EP2419Y] (AB76409), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PICC/IFIPWBFlow Cyt (Intra)
Human
Tested
Not recommended
Tested
Tested
Tested

Tested
Tested

Species

Human

Dilution info

1/100 - 1/250

Notes

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

Not recommended
Not recommended

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/20

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000 - 1/2000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/20

Notes

Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Associated Products

Select an associated product type

12 products for Alternative Product

Target data

Function

Key transcriptional regulator of type I interferon (IFN)-dependent immune responses which plays a critical role in the innate immune response against DNA and RNA viruses (PubMed:22394562, PubMed:25636800, PubMed:27302953). Regulates the transcription of type I IFN genes (IFN-alpha and IFN-beta) and IFN-stimulated genes (ISG) by binding to an interferon-stimulated response element (ISRE) in their promoters (PubMed:11846977, PubMed:16846591, PubMed:16979567, PubMed:20049431, PubMed:32972995). Acts as a more potent activator of the IFN-beta (IFNB) gene than the IFN-alpha (IFNA) gene and plays a critical role in both the early and late phases of the IFNA/B gene induction (PubMed:16846591, PubMed:16979567, PubMed:20049431). Found in an inactive form in the cytoplasm of uninfected cells and following viral infection, double-stranded RNA (dsRNA), or toll-like receptor (TLR) signaling, is phosphorylated by IKBKE and TBK1 kinases (PubMed:22394562, PubMed:25636800, PubMed:27302953). This induces a conformational change, leading to its dimerization and nuclear localization and association with CREB binding protein (CREBBP) to form dsRNA-activated factor 1 (DRAF1), a complex which activates the transcription of the type I IFN and ISG genes (PubMed:16154084, PubMed:27302953, PubMed:33440148). Can activate distinct gene expression programs in macrophages and can induce significant apoptosis in primary macrophages (PubMed:16846591). In response to Sendai virus infection, is recruited by TOMM70:HSP90AA1 to mitochondrion and forms an apoptosis complex TOMM70:HSP90AA1:IRF3:BAX inducing apoptosis (PubMed:25609812). Key transcription factor regulating the IFN response during SARS-CoV-2 infection (PubMed:33440148).

Alternative names

Interferon regulatory factor 3, IRF-3, IRF3

Recommended products

Rabbit Recombinant Monoclonal IRF3 antibody. Suitable for IHC-P, IP, WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 25 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EP2419Y

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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.

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 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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

IRF3 also known as Interferon Regulatory Factor 3 acts as an important transcription factor in the immune response. It has a molecular weight of approximately 47 kDa. The IRF3 protein is mainly expressed in the cytoplasm and nucleus of various cell types including immune cells such as macrophages and dendritic cells. The protein becomes activated through phosphorylation a process frequently identified in its phosphorylated form phospo-IRF3 or p-IRF3 which facilitates its role in immune function.

Biological function summary

IRF3 participates in the regulation of type I interferon (IFN) response a fundamental antiviral defense mechanism. IRF3 when phosphorylated forms a complex with CBP/p300 which then translocates to the nucleus to drive the expression of IFN-stimulated genes. This action strengthens the innate immune response and boosts the body's ability to counteract viral infections. Its activity and regulation are significant for maintaining a balanced immune response without excessive inflammation.

Pathways

IRF3 is involved in the Toll-like receptor (TLR) and RIG-I-like receptor (RLR) signaling pathways both essential in pathogen recognition and response. Within these pathways IRF3 interacts with proteins such as MAVS and TBK1 to propagate immune signaling. The activation of IRF3 in these pathways results in the production of type I interferons and other cytokines orchestrating an effective antiviral response. These interactions highlight the protein's central role in mediating immune signaling cascades.

Associated diseases and disorders

IRF3's malfunction or deregulation can contribute to autoimmune diseases and antiviral deficiencies. Conditions such as systemic lupus erythematosus (SLE) and chronic hepatitis B infection are linked to IRF3 activity. During autoimmune responses or viral persistence the aberrant activation of IRF3 can lead to inappropriate immune responses. The connection with proteins like STAT1 in these conditions highlights the complex network IRF3 engages in facilitating its impact on disease progression and immune dysregulation.

Product promise

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7 product images

  • Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409), expandable thumbnail

    Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409)

    Lanes 1 - 2: Merged signal (red and green). Green - ab76409 observed at 50 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab76409 was shown to react with IRF3 in wild-type A549 cells in western blot with loss of signal observed in IRF3 knockout cell line Human IRF3 knockout A549 cell line ab267097 (IRF3 knockout cell lysate Human IRF3 knockout A549 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 ab76409 and Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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.

    All lanes: Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409) at 1/1000 dilution

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: IRF3 knockout A549 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 47 kDa

    Observed band size: 50 kDa

  • Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409), expandable thumbnail

    Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409)

    We are unsure how to define the extra bands.

    All lanes: Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409) at 1/1000 dilution

    Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

    Lane 3: U-937 (Human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

    Lane 4: THP-1 (Human monocytic leukemia monocyte) whole cell lysate at 20 µg

    Lane 5: Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 47 kDa

  • Immunoprecipitation - Anti-IRF3 antibody [EP2419Y] (ab76409), expandable thumbnail

    Immunoprecipitation - Anti-IRF3 antibody [EP2419Y] (ab76409)

    Purified ab76409 at 1/20 dilution (0.8μg) immunoprecipitating IRF3 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
    Lane 2 (+): ab76409 + HeLa whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab76409 in HeLa whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 47 kDa
    Fresh lysate need to be used to avoid protein degradation.

    All lanes: Immunoprecipitation - Anti-IRF3 antibody [EP2419Y] (ab76409)

    Predicted band size: 47 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EP2419Y] (ab76409), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EP2419Y] (ab76409)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue sections labeling IRF3 with Purified 76409 at 1:100 dilution (1.55 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409), expandable thumbnail

    Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409)

    Lanes 1 - 4: Merged signal (red and green). Green - ab76409 observed at 50 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.

    ab76409 was shown to react with IRF3 in wild-type HeLa. Loss of signal was observed when knockout cell line Human IRF3 knockout HeLa cell line ab255345 (knockout cell lysate Human IRF3 knockout HeLa cell lysate ab263784) was used. Wild-type and IRF3 knockout samples were subjected to SDS-PAGE. ab76409 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.

    All lanes: Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409) at 1/1000 dilution

    Lane 1: Jurkat cell lysate at 20 µg

    Lane 2: MCF7 cell lysate at 20 µg

    Lane 3: Wild-type HeLa cell lysate at 20 µg

    Lane 4: IRF3 knockout HeLa cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 47 kDa

    Observed band size: 47 kDa, 50 kDa

  • Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409), expandable thumbnail

    Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409)

    Lanes 1 - 4: Merged signal (red and green). Green - ab76409 observed at 50 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - 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 Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.

    All lanes: Western blot - Anti-IRF3 antibody [EP2419Y] (ab76409) at 1/1000 dilution

    Lane 1: Wild-type HAP1 cell lysate at 20 µg

    Lane 2: IRF3 knockout HAP1 cell lysate at 20 µg

    Lane 3: HeLa cell lysate at 20 µg

    Lane 4: Jurkat cell lysate at 20 µg

    Predicted band size: 47 kDa

  • Flow Cytometry (Intracellular) - Anti-IRF3 antibody [EP2419Y] (ab76409), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-IRF3 antibody [EP2419Y] (ab76409)

    Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling IRF3 with Purified 76409 at 1/20 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

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