Product nameAnti-IRF3 antibody [EP2419Y]
See all IRF3 primary antibodies
DescriptionRabbit monoclonal [EP2419Y] to IRF3
Tested applicationsSuitable for: WB, IP, IHC-P, Flow Cytmore details
Unsuitable for: ICC
Species reactivityReacts with: Human
Synthetic peptide within Human IRF3 (C terminal). The exact sequence is proprietary.
- U937, HeLa, MCF7 and Jurkat cell lysates; human tonsil tissue.
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 rabbit monoclonal antibody.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
PurityTissue culture supernatant
Our Abpromise guarantee covers the use of ab76409 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/2000. Predicted molecular weight: 47 kDa.|
|IHC-P||1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionMediates 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 specificityExpressed constitutively in a variety of tissues.
Sequence similaritiesBelongs to the IRF family.
Contains 1 IRF tryptophan pentad repeat DNA-binding domain.
modificationsConstitutively 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 localizationCytoplasm. 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
- IIAE7 antibody
- Interferon regulatory factor 3 antibody
- IRF 3 antibody
Lane 1: Jurkat cell lysate (20 µg)
Lane 2: MCF7 cell lysate (20 µg)
Lane 3: HeLa wildtype cell lysate (20 µg)
Lane 4: IRF3 HeLa knockout cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab76409 observed at 47 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab76409 was shown to react with IRF3 in HeLa wildtype. Loss of signal was observed when knockout sample ab263784 was used. Wild-type and IRF3 knockout samples were subjected to SDS-PAGE. ab76409 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.
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.
All lanes : Anti-IRF3 antibody [EP2419Y] (ab76409) at 1/2000 dilution
Lane 1 : U937 cell lysate
Lane 2 : Hela cell lysate
Lane 3 : MCF7 cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 47 kDa
Observed band size: 47 kDa
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.
Flow cytometric analysis of permeabilized U937 cells using anti- IRF-3 RabMAb (red) (ab76409) or a rabbit IgG (negative) (green)
This product has been referenced in:
- Tajpara P et al. A Preclinical Model for Studying Herpes Simplex Virus Infection. J Invest Dermatol 139:673-682 (2019). Read more (PubMed: 30414908) »
- Reisländer T et al. BRCA2 abrogation triggers innate immune responses potentiated by treatment with PARP inhibitors. Nat Commun 10:3143 (2019). Read more (PubMed: 31316060) »