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Products:Immunology >> Innate Immunity >> Cytokines >> Interferons
Overview
Product name
Anti-IRF3 antibody
See all IRF3 products (14) ...
Description
Rabbit polyclonal to IRF3
Tested applications
WB, ICC/IF, IHC-Pmore details
Cross reactivity
Reacts with
Human
Predicted to work with
Chimpanzee
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Human IRF3.
(Peptide available as ab87370.)
Positive control
This antibody gave a positive signal in HeLa, Jurkat and THP1 whole cell lysates.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Immunology >> Innate Immunity >> TLR Signaling
Epigenetics and Nuclear Signaling >> Transcription >> Other factors
Immunology >> Innate Immunity >> Cytokines >> Interferons
Applications
Show applications key
Our Abpromise guarantee covers the use of ab76367 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use a concentration of 1 µg/mlDetects a band of approximately 50 kDa (predicted molecular weight: 47 kDa).
ICC/IF: Use a concentration of 5 µg/ml
IHC-P: Use a concentration of 5 µg/mlPerform heat mediated antigen retrieval before commencing with IHC staining protocol.
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 accessionQ14653
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Interferon regulatory factor 3 antibody
- IRF 3 antibody
- IRF-3 antibody
see all
Database links
- Entrez Gene: 3661 Human
- GenBank: AAO24761 Human
- Omim: 603734 Human
- SwissProt: Q14653 Human
- SwissProt: Q5FBY1 Human
- SwissProt: Q5FBY2 Human
- SwissProt: Q5FBY4 Human
- Unigene: 289052 Human
see all
Anti-IRF3 antibody images:
Western blot - IRF3 antibody (ab76367)
All lanes : Anti-IRF3 antibody (ab76367) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lane 3 : THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 47 kDa
Observed band size : 50 kDa (why is the actual band size different from the predicted?)
Additional bands at : 30 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 5 minutes
IRF3 contains a potential phosphorylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted.
Immunocytochemistry/ Immunofluorescence - IRF3 antibody (ab76367)
ICC/IF image of ab76367 stained HeLa cells. The cells were 4% PFA 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 (ab76367, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit 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. This antibody also gave a positive result in 4% PFA fixed (10 min) MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - IRF3 antibody (ab76367)
IHC image of IRF3 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab76367, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
References for Anti-IRF3 antibody (ab76367)
ab76367 has not yet been referenced specifically in any publications.
Publishing research using ab76367? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"