[KO Validated] IRF3 Rabbit mAb - Q14653

Core Product Specifications and Parameters

Biological Background: IRF3 Function and Localization

• IRF3 is a key transcriptional regulator of type I interferon (IFN)-dependent immune responses, playing a critical role in innate immunity against DNA and RNA viruses. It regulates the transcription of type I IFN genes (IFN-alpha and IFN-beta) and IFN-stimulated genes (ISG) by binding to interferon-stimulated response elements (ISRE) in their promoters. Related references: PMID:22394562 PMID:24049179 PMID:25636800 PMID:11846977 PMID:16846591 PMID:16979567
• In uninfected cells, IRF3 resides in the cytoplasm in an inactive form. Upon viral infection, double-stranded RNA (dsRNA), or toll-like receptor (TLR) signaling, it is phosphorylated by IKBKE and TBK1 kinases, inducing a conformational change that leads to dimerization, nuclear localization, and association with CREBBP to form the dsRNA-activated factor 1 (DRAF1) complex, which activates type I IFN and ISG genes. Related references: PMID:22394562 PMID:25636800 PMID:27302953 PMID:16154084 PMID:33440148
• IRF3 can activate distinct gene expression programs in macrophages and induce significant apoptosis in primary macrophages. In response to Sendai virus infection, IRF3 is recruited by TOMM70:HSP90AA1 to mitochondria, where it forms an apoptosis complex TOMM70:HSP90AA1:IRF3:BAX, inducing apoptosis. Related references: PMID:16846591 PMID:25609812
• IRF3 is a key transcription factor regulating the IFN response during SARS-CoV-2 infection. Related references: PMID:33440148
• Subcellular localization includes cytoplasm, nucleus, and mitochondrion, consistent with its dynamic regulation during signaling.
• Post-translational modifications include phosphorylation (critical for activation), acetylation, disulfide bond formation, isopeptide bond, and Ubl conjugation.
• Expressed constitutively in a variety of tissues, underscoring its broad role in antiviral defense.
• Alternative splicing produces multiple isoforms, potentially modulating its function in different contexts.

Experimental Guidance and Technical Tips

• The immunogen corresponds to amino acids 1-260 of human IRF3 (NP_001184051.1), covering the N-terminal region which includes DNA-binding and regulatory domains. This design may detect both unphosphorylated and phosphorylated forms, making it suitable for monitoring total IRF3 levels.
• For Western blotting, consider using lysates from cells stimulated with viral mimics (e.g., poly(I:C)) or interferons to observe activation-dependent band shifts indicative of phosphorylation. Validate in the relevant sample system, such as virus-infected or TLR-activated cells.
• In immunohistochemistry and immunofluorescence, antigen retrieval and appropriate fixation methods should be optimized to preserve the native conformation of IRF3, especially as it undergoes phosphorylation-dependent nuclear translocation.
• For immunoprecipitation, ensure gentle lysis conditions to maintain protein-protein interactions, as IRF3 forms complexes with partners like CREBBP and TBK1.
• The antibody’s reactivity with mouse and rat species (based on sequence homology) allows cross-species studies, but it is recommended to verify reactivity in the specific experimental model.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio provides diagnostic manufacturers, laboratories, and research institutes with one-stop access to IVD raw materials, technical services, and consulting, covering every stage from concept to clinic. As a trusted partner in immune-oncology and infectious disease research, we offer validated antibody pairs, optimized monoclonal and polyclonal antibodies, bulk ancillary reagents, and support for rare target raw-material sourcing. With this IRF3 monoclonal antibody, we empower your innate immunity and antiviral response studies, ensuring reliable supply for your diagnostic and research needs.