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Immunofluorescence (IF)/Immunocytochemistry (ICC) (Staining HepG2 cells(4h of LPS treatment) by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25 degree C. Samples were then incubated with primary Ab(AF7456 1:200) and mouse anti-beta tubulin Ab(T0023 1:200) for 1 hour at 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(Green) were used as the secondary antibody.The nuclear counter stain is DAPI(blue).)

Rabbit IRF3 Polyclonal Antibody | anti-IRF3 antibody

Phospho-IRF3 (Ser396) Antibody

Reactivity
Human, Mouse, Rat
Applications
Immunofluorescence, Immunocytochemistry, ELISA
Purity
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Synonyms
IRF3; Polyclonal Antibody; Phospho-IRF3 (Ser396) Antibody; IIAE7; Interferon regulatory factor 3; IRF 3; IRF-3; IRF3_HUMAN; MGC94729; anti-IRF3 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Phospho-IRF3 (Ser396) Antibody detects endogenous levels of IRF3 only when phosphorylated at Ser396.
Tissue Specificity: Expressed constitutively in a variety of tissues.
Purity/Purification
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Form/Format
Liquid. Rabbit IgG in phosphate buffered saline, pH7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Concentration
1mg/ml (varies by lot)
Applicable Applications for anti-IRF3 antibody
Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
Application Notes
IF/ICC: 1:100-1:500
Peptide ELISA: 1:20,000-1:40,000
Immunogen
A synthesized peptide derived from human IRF3 around the phosphorylation site of Ser396.
Conjugation
Unconjugated
Fragment
Fab fragment
Post Translational Modifications
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.
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.
Similarity
Belongs to the IRF family.
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).
Preparation and Storage
Store at -20 degree C. Stable for 12 months from date of receipt.

Immunofluorescence (IF)/Immunocytochemistry (ICC)

(Staining HepG2 cells(4h of LPS treatment) by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25 degree C. Samples were then incubated with primary Ab(AF7456 1:200) and mouse anti-beta tubulin Ab(T0023 1:200) for 1 hour at 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(Green) were used as the secondary antibody.The nuclear counter stain is DAPI(blue).)

Immunofluorescence (IF)/Immunocytochemistry (ICC) (Staining HepG2 cells(4h of LPS treatment) by IF/ICC. The samples were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25 degree C. Samples were then incubated with primary Ab(AF7456 1:200) and mouse anti-beta tubulin Ab(T0023 1:200) for 1 hour at 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(Green) were used as the secondary antibody.The nuclear counter stain is DAPI(blue).)
Related Product Information for anti-IRF3 antibody
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.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
47,219 Da
NCBI Official Full Name
interferon regulatory factor 3 isoform 4
NCBI Official Synonym Full Names
interferon regulatory factor 3
NCBI Official Symbol
IRF3
NCBI Protein Information
interferon regulatory factor 3
UniProt Protein Name
Interferon regulatory factor 3
UniProt Gene Name
IRF3
UniProt Synonym Gene Names
IRF-3
UniProt Entry Name
IRF3_HUMAN

NCBI Description

This gene encodes a member of the interferon regulatory transcription factor (IRF) family. The encoded protein is found in an inactive cytoplasmic form that upon serine/threonine phosphorylation forms a complex with CREBBP. This complex translocates to the nucleus and activates the transcription of interferons alpha and beta, as well as other interferon-induced genes. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene. [provided by RefSeq, Nov 2011]

Uniprot Description

IRF3: interferon regulatory factor 3, a member of the interferon regulatory transcription factor (IRF) family. IRF3 is found in an inactive cytoplasmic form that upon serine/threonine phosphorylation forms a complex with CREBBP. This complex translocates to the nucleus and activates the transcription of interferons alpha and beta, as well as other interferon-induced genes.

Protein type: Transcription factor; DNA-binding

Chromosomal Location of Human Ortholog: 19q13.3-q13.4

Cellular Component: nucleoplasm; cytoplasm; cytosol; nucleus

Molecular Function: identical protein binding; protein binding; protein homodimerization activity; DNA binding; transcription cofactor activity; transcription factor activity

Biological Process: negative regulation of interferon-beta biosynthetic process; transcription from RNA polymerase II promoter; positive regulation of I-kappaB kinase/NF-kappaB cascade; viral reproduction; apoptosis; cytokine and chemokine mediated signaling pathway; MyD88-independent toll-like receptor signaling pathway; negative regulation of transcription from RNA polymerase II promoter; toll-like receptor 3 signaling pathway; negative regulation of defense response to virus by host; response to exogenous dsRNA; positive regulation of interferon type I production; lipopolysaccharide-mediated signaling pathway; positive regulation of interferon-beta production; toll-like receptor signaling pathway; innate immune response; interferon type I biosynthetic process; toll-like receptor 4 signaling pathway; response to DNA damage stimulus; negative regulation of interferon type I production; defense response to virus; positive regulation of interferon-alpha production; positive regulation of cytokine secretion

Disease: Herpes Simplex Encephalitis, Susceptibility To, 7

Research Articles on IRF3

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Product Notes

The IRF3 irf3 (Catalog #AAA9613999) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Phospho-IRF3 (Ser396) Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's IRF3 can be used in a range of immunoassay formats including, but not limited to, Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA). IF/ICC: 1:100-1:500 Peptide ELISA: 1:20,000-1:40,000. Researchers should empirically determine the suitability of the IRF3 irf3 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "IRF3, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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