Revision 1

#45011Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

M

UniProt ID:

#P70434

Entrez-Gene Id:

#54123

Cell Signaling Technology

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
IRF-7 Rabbit mAb Coated Microwells 53117 96 tests +4C
IRF-7 Rabbit Detection mAb 85662 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 Total IRF-7 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IRF-7 protein 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 IRF-7 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 IRF-7 protein. Learn more about your ELISA kit options here.

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

Specificity/Sensitivity

The PathScan® RP Total IRF-7 Sandwich ELISA Kit detects endogenous levels of IRF-7 protein. 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-7, which is functionally similar to IRF-3, is preferentially expressed in lymphoid cells and induced by virus, LPS, and IFN-α (3-5). IRF-7 plays an essential role in the induction of type I interferon in response to viral infection (6-8). IRF-7 is regulated at multiple serine phosphorylation sites near the carboxyl terminus, similar to IRF-3, which are required for nuclear translocation, DNA binding, and transcriptional activity (9-11).

  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. Au, W.C. et al. (1998) J Biol Chem 273, 29210-7.
  4. Wathelet, M.G. et al. (1998) Mol Cell 1, 507-18.
  5. Marié, I. et al. (1998) EMBO J 17, 6660-9.
  6. Sato, M. et al. (2000) Immunity 13, 539-48.
  7. Honda, K. et al. (2005) Nature 434, 772-7.
  8. Colina, R. et al. (2008) Nature 452, 323-8.
  9. Lin, R. et al. (2000) J Biol Chem 275, 34320-7.
  10. Yang, H. et al. (2003) J Biol Chem 278, 15495-504.
  11. Caillaud, A. et al. (2005) J Biol Chem 280, 17671-7.

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 1
    #45011

    PathScan® RP Total IRF-7 Sandwich ELISA Kit

    PathScan® RP Total IRF-7 Sandwich ELISA Kit: Image 1 Expand Image
    Figure 1. Co-transfection of 293T cells with constructs expressing mouse IRF-7 and human IKKε increases phosphorylation of IRF-7 at Ser437/438 compared to cells transfected with only an IRF-7 expression construct, but does not affect the level of total IRF-7 protein. IRF-7 protein is also expressed at higher levels in 293T cells transfected with an IRF-7 expression construct compared to mock transfected cells. The relationship between lysate protein concentration from IRF-7/IKKε co-transfected, IRF-7-only transfected, and mock transfected 293T cells and the absorbance at 450 nm using the PathScan® RP Total IRF-7 Sandwich ELISA Kit #45011 is shown in the upper figure. The corresponding western blots using mouse IRF-7 antibody (left panel), mouse phospho-IRF-7 (Ser437/438) antibody (middle panel), and β-Actin antibody (right panel) are shown in the lower figure. 293T cells were either co-transfected with constructs expressing Myc-DDK-tagged mouse IRF-7 (mIRF-7) and GST-tagged human IKKε (hIKKε), transfected with a construct expressing Myc-DDK-tagged mouse IRF-7, or mock transfected, and then lysed.