Key features and details
- Rabbit polyclonal to IRF3
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Mouse, Human
- Isotype: IgG
Product nameAnti-IRF3 antibody
See all IRF3 primary antibodies
DescriptionRabbit polyclonal to IRF3
Specificityab25950 recognises IRF3.
Tested applicationsSuitable for: ICC/IF, IHC-P, WBmore details
Species reactivityReacts with: Mouse, Human
A synthetic peptide corresponding to 14 amino acids near the carboxyterminus of IRF3 (Human)
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Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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Storage instructionsShipped 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 bufferpH: 7.2
Preservative: 0.02% Sodium azide
Constituent: 99% PBS
Concentration information loading...
PurityImmunogen affinity purified
Purification notesIRF3 Antibody is affinity chromatography purified via peptide column.
Our Abpromise guarantee covers the use of ab25950 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 44 kDa (predicted molecular weight: 47 kDa).|
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 : Anti-IRF3 antibody (ab25950) at 1 µg/ml
Lane 2 : Anti-IRF3 antibody (ab25950) at 2 µg/ml
Lane 3 : Anti-IRF3 antibody (ab25950) at 4 µg/ml
All lanes : Ramos whole cell lysate.
Predicted band size: 47 kDa
Observed band size: ~44 kDa why is the actual band size different from the predicted?
ab25950 stained MCF-7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab25950 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Anti-IRF3 antibody (ab25950) at 1/1000 dilution + HT-29 at 30 µg
HRP conjugated Anti-rabbit IgG, Goat polyclonal at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 47 kDa
Samples were incubated with primary antibody for 18 hours at 4ºC and blocked with 5% BSA for 1 hour at room temperature.
Immunofluorescence of IRF3 in Mouse Kidney cells using ab25950 at 20 ug/ml.
ab25950 at 2µg/ml staining IRF3 in mouse kidney by IHC.
ab25950 has been referenced in 25 publications.
- Yu H et al. Association of the TBK1 mutation p.Ile334Thr with frontotemporal dementia and literature review. Mol Genet Genomic Med 7:e547 (2019). PubMed: 30672142
- Lee HC et al. Released Tryptophanyl-tRNA Synthetase Stimulates Innate Immune Responses against Viral Infection. J Virol 93:N/A (2019). PubMed: 30355684
- Kim JH et al. Small Heterodimer Partner Controls the Virus-Mediated Antiviral Immune Response by Targeting CREB-Binding Protein in the Nucleus. Cell Rep 27:2105-2118.e5 (2019). PubMed: 31091449
- Liu TM et al. Transcription Factor MafB Suppresses Type I Interferon Production by CD14+ Monocytes in Patients With Chronic Hepatitis C. Front Microbiol 10:1814 (2019). PubMed: 31447817
- DeDiego ML et al. Interferon-Induced Protein 44 Interacts with Cellular FK506-Binding Protein 5, Negatively Regulates Host Antiviral Responses, and Supports Virus Replication. mBio 10:N/A (2019). PubMed: 31455651