Revision 8

#54395Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

M

UniProt ID:

#P70671

Entrez-Gene Id:

#54131

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
Phospho-IRF-3 (Ser379) Rabbit mAb Coated Microwells 20719 96 tests +4C
IRF-3 Rabbit Detection mAb 39239 1 ea Red (Lyophilized) +4C
HRP Diluent 13515 5.5 ml Red +4C
TMB Substrate 7004 11 ml +4C
STOP Solution 7002 11 ml +4C
Sealing Tape 54503 2 ea +4C
ELISA Wash Buffer (20X) 9801 25 ml +4C
Cell Lysis Buffer (10X) 9803 15 ml -20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The rapid protocol (RP) PathScan® RP Phospho-IRF-3 (Ser379) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IRF-3 protein phosphorylated at Ser379 in a reduced assay time of 1.5 hours. Incubation of cell lysates and detection antibody on the coated microwell plate forms a sandwich with phospho-IRF-3 (Ser379) in a single step. The plate is then extensively washed and TMB reagent is added for signal development. The magnitude of absorbance for the developed color is proportional to the quantity of phospho-IRF-3 (Ser379). Learn more about all of your ELISA kit options here.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

The PathScan® RP Phospho-IRF-3 (Ser379) Sandwich ELISA Kit detects endogenous levels of mouse IRF-3 protein phosphorylated at Ser379 (equivalent to Ser386 of human IRF-3). The kit sensitivity is shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

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, IRF-9/ISGF3γ, 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 carboxyl-terminal serine and threonine 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 of 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).

  1. Taniguchi, T. et al. (2001) Annu Rev Immunol 19, 623-55.
  2. Honda, K. and Taniguchi, T. (2006) Nat Rev Immunol 6, 644-58.
  3. Hiscott, J. et al. (1999) J Interferon Cytokine Res 19, 1-13.
  4. Kim, T.Y. et al. (2003) J Biol Chem 278, 15272-8.
  5. Yoneyama, M. et al. (2002) J Interferon Cytokine Res 22, 73-6.
  6. Fitzgerald, K.A. et al. (2003) Nat Immunol 4, 491-6.
  7. Kopp, E. and Medzhitov, R. (2003) Curr Opin Immunol 15, 396-401.
  8. Servant, M.J. et al. (2003) J Biol Chem 278, 9441-7.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PathScan is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 8
    #54395

    PathScan® RP Phospho-IRF-3 (Ser379) Sandwich ELISA Kit

    PathScan® RP Phospho-IRF-3 (Ser379) Sandwich ELISA Kit: Image 1 Expand Image
    Figure 1. Treatment of J774A.1 cells with 2'3'c-GAMP stimulates phosphorylation of IRF-3 at Ser379. The relationship between lysate protein concentration from untreated and 2'3'c-GAMP-treated J774A.1 cells and the absorbance at 450 nm using the PathScan® RP Phospho-IRF-3 (Ser379) Sandwich ELISA Kit #54395 is shown in the upper figure. The corresponding western blots using phospho-IRF-3 (Ser379) antibody (left panel) and IRF-3 antibody (right panel) are shown in the lower figure. J774A.1 cells were treated with 2'3'c-GAMP (10 µg/mL) for 6 hours at 37°C and then lysed.