Preparation And Storage
Recommended Assay Procedures
Clone SL-12.1 can be used for western blot analysis (0.5-2.0 µg/ml). Other reported applications include immunoprecipitation and immunofluorescence microscopy. The antibody will recognize both the nuclear and cytoplasmic forms of IRF-3 and can block IRF-3 DNA binding, although this application has not been tested at BD Biosciences. Jurkat cells are recommended as a positive control (Cat. No. 611451).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
Companion Products
The SL-12.1 monoclonal antibody specifically recognizes human Interferon regulatory factor 3 (IRF-3). Viral infection in mammals can lead to the induction of multiple pathways as part of the host defense mechanism. One of the major pathways activated is the JAK-STAT pathway by various interferons (IFNα and IFNβ). These IFNs exert their influence via transcriptional activation of specific target genes involved in antiviral defense, for example the chemokine ISG15 gene or the major histocompatibility complex class I and II molecules. These genes in turn are regulated by the JAK-STAT signaling pathway and through interferon regulatory factors (IRFs). IRFs are a family of transcription factors that possess a broad range of activities. IRF-3 is one of nine members which all share a common DNA binding domain which binds to an IFN stimulated response element (ISRE) found in the majority of IFN-inducible promoters. The IRF3 gene expresses a 50 kDa protein which is constitutively expressed in all tissues. The protein undergoes post-translational modification as well as dimerization and is translocated from the cytoplasm to the nucleus upon viral infection or exposure to dsRNA.
Development References (8)
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Au WC, Moore PA, Lowther W, Juang YT, Pitha PM. Identification of a member of the interferon regulatory factor family that binds to the interferon-stimulated response element and activates expression of interferon-induced genes. Proc Natl Acad Sci U S A. 1995; 92(25):11657-11661. (Biology). View Reference
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Darnell JE Jr, Kerr IM, Stark GR. Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins. Science. 1994; 264(5164):1415-1421. (Biology). View Reference
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Hiscott J, Pitha P, Genin P, et al. Triggering the interferon response: the role of IRF-3 transcription factor.. J Interferon Cytokine Res. 1999; 19(1):1-13. (Biology). View Reference
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Loh JE, Chang CH, Fodor WL, Flavell RA. Dissection of the interferon gamma-MHC class II signal transduction pathway reveals that type I and type II interferon systems share common signalling component(s). EMBO J. 1992; 11(4):1351-1363. (Biology). View Reference
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Mamane Y, Heylbroeck C, Genin P, et al. Interferon regulatory factors: the next generation. Gene. 1999; 237(1):1-14. (Biology). View Reference
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Reich N, Evans B, Levy D, Fahey D, Knight E Jr, Darnell JE Jr. Interferon-induced transcription of a gene encoding a 15-kDa protein depends on an upstream enhancer element. Proc Natl Acad Sci U S A. 1987; 84(18):6394-6398. (Biology). View Reference
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Ronco LV, Karpova AY, Vidal M, Howley PM. Human papillomavirus 16 E6 oncoprotein binds to interferon regulatory factor-3 and inhibits its transcriptional activity. Genes Dev. 1998; 12(13):2061-2072. (Clone-specific: Western blot). View Reference
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Wathelet MG, Lin CH, Parekh BS, Ronco LV, Howley PM, Maniatis T. Virus infection induces the assembly of coordinately activated transcription factors on the IFN-beta enhancer in vivo. Mol Cell. 1998; 1(4):507-518. (Clone-specific: Blocking, Immunofluorescence, Immunoprecipitation, Western blot). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
