Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- HRP Rabbit monoclonal [EPR2418Y] to IRF3
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
- Conjugation: HRP
Product nameHRP Anti-IRF3 antibody [EPR2418Y]
See all IRF3 primary antibodies
DescriptionHRP Rabbit monoclonal [EPR2418Y] to IRF3
Tested applicationsSuitable for: IHC-P, WBmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse
Synthetic peptide within Human IRF3 aa 50-150. The exact sequence is proprietary.
Database link: Q14653
- WB: HeLa, Jurkat and wild-type HAP1 whole cell lysates. IHC-P: Normal human tonsil tissue.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. Store In the Dark.
Storage bufferpH: 7.40
Preservative: 0.1% 10% Proclin 300 Solution
Constituents: PBS, 30% Glycerol, 1% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab205443 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.|
|WB||1/2000. Detects a band of approximately 51 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
All lanes : HRP Anti-IRF3 antibody [EPR2418Y] (ab205443) at 1/2000 dilution
Lane 1 :
HeLa whole cell lysate (ab150035) at 10 µg
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg
Lane 3 : Wild-type HAP1 cell lysate at 20 µg
Lane 4 : IRF3 knockout HAP1 cell lysate at 20 µg
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 47 kDa
Observed band size: 51 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutes
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab205443 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
IHC image of IRF3 staining in a section of formalin-fixed paraffin-embedded normal human tonsil tissue*, performed on a Leica BONDTM. The section was pre-treated using heat mediated antigen retrieval with TRIS/EDTA (pH9, epitope retrieval solution 2) for 20mins. The section was then incubated with ab205443, 1/50 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
ab205443 has not yet been referenced specifically in any publications.