Anti-IRF3 antibody [IRF35I218] (ab50772)
Key features and details
- Mouse monoclonal [IRF35I218] to IRF3
- Suitable for: ICC/IF, WB, Flow Cyt
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-IRF3 antibody [IRF35I218]
See all IRF3 primary antibodies -
Description
Mouse monoclonal [IRF35I218] to IRF3 -
Host species
Mouse -
Tested applications
Suitable for: ICC/IF, WB, Flow Cytmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment: VAWVNKSRTR FRIPWKHGLR QDAQQEDFGI FQAWAEATGA YVPGRDKPDL PTWKRNFRSA LNRKEGLRLA EDRSKD, corresponding to amino acids 24-150 of Human IRF3
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Positive control
- HEK293 whole cell lysate RIPA lysate of HeLa cells Hela cells
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. -
Storage buffer
pH: 7.40
Preservative: 0.05% Sodium azide
Constituents: 0.316% Tris HCl, 1% BSA, 1.16% Sodium chloride -
Concentration information loading...
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Purity
Tissue culture supernatant -
Purification notes
Concentrated and dialyzed culture medium (protein free medium) of hybridoma -
Clonality
Monoclonal -
Clone number
IRF35I218 -
Isotype
IgG1 -
Research areas
Associated products
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Assay kits
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab50772 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ICC/IF |
1/10.
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WB | (3) |
1/50 - 1/100. Detects a band of approximately 55 kDa (predicted molecular weight: 47 kDa).
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Flow Cyt |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Notes |
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ICC/IF
1/10. |
WB
1/50 - 1/100. Detects a band of approximately 55 kDa (predicted molecular weight: 47 kDa). |
Flow Cyt
Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Target
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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
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 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. - Information by UniProt
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Database links
- Entrez Gene: 3661 Human
- Omim: 603734 Human
- SwissProt: Q14653 Human
- Unigene: 289052 Human
- Unigene: 75254 Human
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Alternative names
- IIAE7 antibody
- Interferon regulatory factor 3 antibody
- IRF 3 antibody
see all
Images
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Anti-IRF3 antibody [IRF35I218] (ab50772) at 1/50 dilution + HT-1080 whole cell lysate at 25 µg
Secondary
Anti mouse IgG at 1/2500 dilution
Predicted band size: 47 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted? -
Overlay histogram showing HeLa cells stained with ab50772 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab50772, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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Immunofluorescent staining of HeLa cells with a 1/10 dilution of ab50772 as primary antibody and AlexaFluor®488 conjugated Goat Anti-mouse IgG diluted 1/200 as the secondary antibody
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Anti-IRF3 antibody [IRF35I218] (ab50772) at 1/100 dilution + Ripa lysate of Hela cells at 500 µg
Secondary
Anti-mouse IgG at 1/1000 dilution
Predicted band size: 47 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?
IRF3 antibody [IRF35I218] (ab50772) at 1/100 dilution + Ripa lysate of Hela cells at 500 µg Secondary Anti-mouse IgG at 1/1000 dilution
Protocols
Datasheets and documents
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Datasheet download
References (9)
ab50772 has been referenced in 9 publications.
- Uhlorn BL et al. Attenuation of cGAS/STING activity during mitosis. Life Sci Alliance 3:N/A (2020). PubMed: 32661021
- Uhlorn BL et al. Vesicular trafficking permits evasion of cGAS/STING surveillance during initial human papillomavirus infection. PLoS Pathog 16:e1009028 (2020). PubMed: 33253291
- Yi L et al. Interferon regulatory factor 3 mediates Poly(I:C)-induced innate immune response and apoptosis in non-small cell lung cancer. Int J Oncol 52:1623-1632 (2018). PubMed: 29512705
- Wang CH et al. Global Screening of Antiviral Genes that Suppress Baculovirus Transgene Expression in Mammalian Cells. Mol Ther Methods Clin Dev 6:194-206 (2017). PubMed: 28831401
- Iqbal J et al. Histone H2B-IFI16 Recognition of Nuclear Herpesviral Genome Induces Cytoplasmic Interferon-ß Responses. PLoS Pathog 12:e1005967 (2016). PubMed: 27764250
- Brownell J et al. Direct, interferon-independent activation of the CXCL10 promoter by NF-?B and interferon regulatory factor 3 during hepatitis C virus infection. J Virol 88:1582-90 (2014). PubMed: 24257594
- Wang J et al. Morphine inhibits murine dendritic cell interleukin 23 production by modulating TLR2 and Nod2 signaling. J Biol Chem : (2011). ICC/IF ; Mouse . PubMed: 21245149
- Holm CK et al. TLR3 ligand polyinosinic:polycytidylic acid induces IL-17A and IL-21 synthesis in human Th cells. J Immunol 183:4422-31 (2009). WB ; Human . PubMed: 19748983
- Chen GY et al. Baculovirus transduction of mesenchymal stem cells triggers the toll-like receptor 3 pathway. J Virol 83:10548-56 (2009). ICC/IF ; Human . PubMed: 19656899