Product Name:
IRF3-pT135
Product Number:
ab-pn610
Target Full Name: Interferon regulatory factor 3
Target Alias: IRF-3
Product Type Specific: IRF3 phosphosite-specific antibody
Antibody Code: PN610
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: T135
Protein UniProt: Q14653
Protein SigNET: IRF3
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human IRF3
Antibody Immunogen Sequence: CSTSD(pT)QEDI
Antibody Immunogen Description: Corresponds to amino acid residues S131 to I139. T135 phosphoryation causes nuclear retention and delayed proteolysis. This is the major in vivo phosphorylation site in IRF3 (≥30 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). IRF3 is known to be phosphorylated at this site in vitro by DNAPK (PRKDC).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 1 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Rhesus macaque.
Scientific Background: IRF3 (Interferon regulatory factor 3) is a 47-50 kDa transcription factor of the the IRF family that mediates innate antiviral responses following viral infection, double-stranded RNA (dsRNA), or toll-like receptor (TLR) signaling. It is found in an autoinhibited, monomeric form in the cytoplasm of resting cells. Upon detection of viral DNA/RNA (via receptors like STING/MAVS), kinases (TBK1/IKKe) phosphorylate IRF3 at Ser-385 and Ser-386, which enables IRF3 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 gene. IRF3 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. IRF3 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. It can activate distinct gene expression programs in macrophages and can induce significant apoptosis in primary macrophages. In response to Sendai virus infection, IRF3 is recruited by TOMM70:HSP90AA1 to mitochondrion and forms an apoptosis complex TOMM70:HSP90AA1:IRF3:BAX inducing apoptosis. It is expressed constitutively in a variety of tissues. Some viruses (e.g., Herpesvirus) inhibit the immune response by inducing the degradation of IRF3 or blocking its translocation. Mutations or dysfunctional regulation of IRF3 are associated with severe infection-induced encephalopathy. This description may include information annotated by UniProt and/or Google AI.