Product: IRF3 Antibody
Catalog: DF6895
Description: Rabbit polyclonal antibody to IRF3
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Dog
Mol.Wt.: 49kDa; 47kD(Calculated).
Uniprot: Q14653
RRID: AB_2838854

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 100ul $280 In stock
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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(88%), Sheep(100%), Dog(100%)
Clonality:
Polyclonal
Specificity:
IRF3 Antibody detects endogenous levels of total IRF3.
RRID:
AB_2838854
Cite Format: Affinity Biosciences Cat# DF6895, RRID:AB_2838854.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

IIAE7; Interferon regulatory factor 3; IRF 3; IRF-3; IRF3; IRF3_HUMAN; MGC94729;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q14653 IRF3_HUMAN:

Expressed constitutively in a variety of tissues.

Description:
Interferon regulatory factors (IRFs) comprise a family of transcription factors that function within the Jak/Stat pathway to regulate interferon (IFN) and IFN-inducible gene expression in response to viral infection (1). IRFs play an important role in pathogen defense, autoimmunity, lymphocyte development, cell growth, and susceptibility to transformation. The IRF family includes nine members: IRF-1, IRF-2, ISGF3γ/p48, IRF-3, IRF-4 (Pip/LSIRF/ICSAT), IRF-5, IRF-6, IRF-7, and IRF-8/ICSBP. All IRF proteins share homology in their amino-terminal DNA-binding domains. IRF family members regulate transcription through interactions with proteins that share similar DNA-binding motifs, such as IFN-stimulated response elements (ISRE), IFN consensus sequences (ICS), and IFN regulatory elements (IRF-E) (2). IRF-3 can inhibit cell growth and plays a critical role in controlling the expression of genes in the innate immune response (1-4). In unstimulated cells, IRF-3 is present in the cytoplasm. Viral infection results in phosphorylation of IRF-3 and leads to its translocation to the nucleus where it activates promoters containing IRF-3-binding sites. Phosphorylation of IRF-3 occurs at a cluster of C-terminal Ser and Thr residues (between 385 and 405), leading to its association with the p300/CBP coactivator protein that promotes DNA binding and transcriptional activity (5). During infection, IRF-3 is likely activated through a pathway that includes activation of Toll-like receptors and a kinase complex that includes IKKε and TBK1 (6,7). IRF-3 is phosphorylated at Ser396 following viral infection, expression of viral nucleocapsid, and double-stranded RNA treatment. These events likely play a role in activation of IRF-3 (8).
Sequence:
MGTPKPRILPWLVSQLDLGQLEGVAWVNKSRTRFRIPWKHGLRQDAQQEDFGIFQAWAEATGAYVPGRDKPDLPTWKRNFRSALNRKEGLRLAEDRSKDPHDPHKIYEFVNSGVGDFSQPDTSPDTNGGGSTSDTQEDILDELLGNMVLAPLPDPGPPSLAVAPEPCPQPLRSPSLDNPTPFPNLGPSENPLKRLLVPGEEWEFEVTAFYRGRQVFQQTISCPEGLRLVGSEVGDRTLPGWPVTLPDPGMSLTDRGVMSYVRHVLSCLGGGLALWRAGQWLWAQRLGHCHTYWAVSEELLPNSGHGPDGEVPKDKEGGVFDLGPFIVDLITFTEGSGRSPRYALWFCVGESWPQDQPWTKRLVMVKVVPTCLRALVEMARVGGASSLENTVDLHISNSHPLSLTSDQYKAYLQDLVEGMDFQGPGES

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Sheep
100
Dog
100
Horse
88
Xenopus
0
Zebrafish
0
Chicken
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q14653 As Substrate

Site PTM Type Enzyme
T3 Phosphorylation
S14 Phosphorylation
K29 Ubiquitination
K70 Sumoylation
K70 Ubiquitination
T75 Phosphorylation
K77 Ubiquitination
K87 Sumoylation
K87 Ubiquitination
S97 Phosphorylation
S123 Phosphorylation
T135 Phosphorylation P78527 (PRKDC)
S173 Phosphorylation P45984 (MAPK9) , P45983 (MAPK8) , Q9UHD2 (TBK1)
S175 Phosphorylation Q9UHD2 (TBK1)
T180 Phosphorylation
S188 Phosphorylation
K193 Ubiquitination
T237 Phosphorylation
T244 Phosphorylation
T253 Phosphorylation
S259 Phosphorylation
K313 Ubiquitination
K315 Ubiquitination
S339 Phosphorylation
T370 Phosphorylation
S385 Phosphorylation Q9UHD2 (TBK1)
S386 Phosphorylation Q14164 (IKBKE) , Q9UQM7 (CAMK2A) , Q9UHD2 (TBK1)
T390 Phosphorylation
S396 Phosphorylation O14920 (IKBKB) , Q14164 (IKBKE) , Q9UHD2 (TBK1)
S398 Phosphorylation O14920 (IKBKB) , Q14164 (IKBKE) , Q9UHD2 (TBK1)
S402 Phosphorylation O14920 (IKBKB) , Q14164 (IKBKE) , Q9UHD2 (TBK1)
T404 Phosphorylation O14920 (IKBKB) , Q9UHD2 (TBK1)
S405 Phosphorylation O14920 (IKBKB) , Q9UHD2 (TBK1)
S427 Phosphorylation

Research Backgrounds

Function:

Key transcriptional regulator of type I interferon (IFN)-dependent immune responses which plays a critical role in the innate immune response against DNA and RNA viruses. Regulates the transcription of type I IFN genes (IFN-alpha and IFN-beta) and IFN-stimulated genes (ISG) by binding to an interferon-stimulated response element (ISRE) in their promoters. Acts as a more potent activator of the IFN-beta (IFNB) gene than the IFN-alpha (IFNA) gene and plays a critical role in both the early and late phases of the IFNA/B gene induction. Found in an inactive form in the cytoplasm of uninfected cells and following viral infection, double-stranded RNA (dsRNA), or toll-like receptor (TLR) signaling, is phosphorylated by IKBKE and TBK1 kinases. This induces a conformational change, leading to its dimerization and nuclear localization and association with CREB binding protein (CREBBP) to form dsRNA-activated factor 1 (DRAF1), a complex which activates the transcription of the type I IFN and ISG genes. Can activate distinct gene expression programs in macrophages and can induce significant apoptosis in primary macrophages.

PTMs:

Constitutively phosphorylated on many Ser/Thr residues. Activated following phosphorylation by TBK1 and IKBKE. Innate adapter protein MAVS, STING1 or TICAM1 are first activated by viral RNA, cytosolic DNA, and bacterial lipopolysaccharide (LPS), respectively, leading to activation of the kinases TBK1 and IKBKE. These kinases then phosphorylate the adapter proteins on the pLxIS motif, leading to recruitment of IRF3, thereby licensing IRF3 for phosphorylation by TBK1. Phosphorylated IRF3 dissociates from the adapter proteins, dimerizes, and then enters the nucleus to induce IFNs.

(Microbial 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.

Subcellular Location:

Cytoplasm. Nucleus.
Note: Shuttles between cytoplasmic and nuclear compartments, with export being the prevailing effect (PubMed:10805757). When activated, IRF3 interaction with CREBBP prevents its export to the cytoplasm (PubMed:10805757).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed constitutively in a variety of tissues.

Subunit Structure:

Monomer. Homodimer; phosphorylation-induced. Interacts (when phosphorylated) with CREBBP. Interacts with MAVS (via phosphorylated pLxIS motif). Interacts with TICAM1 (via phosphorylated pLxIS motif). Interacts with STING1 (via phosphorylated pLxIS motif). Interacts with IKBKE and TBK1. Interacts with TICAM2. Interacts with RBCK1. Interacts with HERC5. Interacts with DDX3X (phosphorylated at 'Ser-102'); the interaction allows the phosphorylation and activation of IRF3 by IKBKE. Interacts with TRIM21 and ULK1, in the presence of TRIM21; this interaction leads to IRF3 degradation by autophagy. Interacts with RIOK3; RIOK3 probably mediates the interaction of TBK1 with IRF3. Interacts with ILRUN; the interaction inhibits IRF3 binding to its DNA consensus sequence. Interacts with LYAR; this interaction impairs IRF3 DNA-binding activity.

(Microbial infection) Interacts with rotavirus A NSP1 (via pLxIS motif); this interaction leads to the proteasome-dependent degradation of IRF3.

(Microbial infection) Interacts with herpes virus 8/HHV-8 protein VIRF1.

(Microbial infection) Interacts with Seneca Valley virus protease 3C; this interaction is involved in the suppression of IRF3 expression and phosphorylation by the virus.

Family&Domains:

Belongs to the IRF family.

Research Fields

· Human Diseases > Infectious diseases: Bacterial > Pertussis.

· Human Diseases > Infectious diseases: Viral > Hepatitis C.

· Human Diseases > Infectious diseases: Viral > Hepatitis B.

· Human Diseases > Infectious diseases: Viral > Measles.

· Human Diseases > Infectious diseases: Viral > Influenza A.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > Herpes simplex infection.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Organismal Systems > Immune system > Toll-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > NOD-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > RIG-I-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Cytosolic DNA-sensing pathway.   (View pathway)

References

1). Commensal cow Roseburia reduces gut-dysbiosis-induced mastitis through inhibiting bacterial translocation by producing butyrate in mice. Cell Reports, 2022 (PubMed: 36417859) [IF=8.8]

2). Zinc Finger CCCH-Type Antiviral Protein 1 Restricts the Viral Replication by Positively Regulating Type I Interferon Response. Frontiers in Microbiology, 2020 (PubMed: 32922375) [IF=5.2]

Application: WB    Species:    Sample: ZC3HAV1 knockdown cells

FIGURE S1 | (A) The positive cells were observed under fluorescent microscope to detect the expression of GFPs. (B) RT-PCR was performed to detect the expression of ZC3HAV1, IFN-β, MxA, TNF, and IL-6 mRNAs in A549 cells expressing sh-ZC3HAV1 or sh-Luc in response to WSN infection. (D) Western blotting was performed to detect the protein expression of IRF3 and p-IRF3 in ZC3HAV1 knockdown cells after the viral infection.

3). Corilagin Interferes With Toll-Like Receptor 3-Mediated Immune Response in Herpes Simplex Encephalitis. Frontiers in Molecular Neuroscience, 2019 (PubMed: 31080403) [IF=4.8]

Application: WB    Species: mouse    Sample: BV-2 cells

Figure 4.| Lentiviral vector-mediated overexpression of TLR3 in BV-2 cells. The lentivirus (10 MOI) was incubated with the BV-2 cells for 72 h. (A) The expression of GFP was observed with a fluorescence microscope after lentivirus was introduced into the BV-2 cells. (B) The mRNA levels of TLR3 and its downstream molecules were measured by RT-PCR. (C) The protein levels were detected by western blotting. Data are shown as the mean ± SD of three independent experiments of triplicate samples.**P < 0.01 for lentivirus-up vs. normal; *P < 0.05 for lentivirus-up vs. normal (by Student’s t-test).

4). RACK1 degrades MAVS to promote bovine ephemeral fever virus replication via upregulating E3 ubiquitin ligase STUB1. VETERINARY MICROBIOLOGY, 2021 (PubMed: 33940459) [IF=3.3]

Application: WB    Species: Mouse    Sample: BHK-21 cell

Fig. 3. RACK1 targets MAVS to inhibit type I IFN signaling pathway. (A) Immunoblot analysis of MAVS, p-TBK1(S172), TBK1, p-IRF3(S386), and IRF3 in negative control and RACK1-overexpressing BHK-21 cell lines infected with BEFV of 0.1MOI for 24 h. Anti-β-actin was used as an internal reference control. (B) HEK-293 T cells were co-transfected with Luc-IFN-β and pRL-TK plasmids plus the indicated expression plasmids together with control or RACK1 plasmids. At 24 h post-transfection, the luciferase activities were measured with the dual-luciferase assay. (C) Immunoblot analysis of MAVS in negative-control and RACK1-overexpressing MDBK cell lines infected with BEFV of 0.1MOI for 24 h. The mean and standard deviation values were determined from three independent experiments; *, P ≤ 0.05.

5). The intervention mechanism of emodin on TLR3 pathway in the process of central nervous system injury caused by herpes virus infection. NEUROLOGICAL RESEARCH, 2021 (PubMed: 33274693) [IF=1.9]

Application: WB    Species: Mice    Sample: brain tissues

Figure 4. The protein expression of TLR3 and its downstream protein in brain tissues of mice.

6). The Reduced Oligomerization of MAVS Mediated by ROS Enhances the Cellular Radioresistance. Oxidative Medicine and Cellular Longevity, 2020 (PubMed: 32190169)

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