Revision 1

#45011

Store at +4C

PathScan® RP Total IRF-7 Sandwich ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
M

UniProt ID:
#P70434

Entrez-Gene Id:
#54123

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Color Storage Temp
IRF-7 Rabbit mAb Coated Microwells5311796 tests+4C
IRF-7 Rabbit Detection mAb856621 eaRed (Lyophilized)+4C
HRP Diluent135155.5 mlRed+4C
TMB Substrate700411 ml+4C
STOP Solution700211 ml+4C
Sealing Tape545032 ea+4C
ELISA Wash Buffer (20X)980125 ml+4C
Cell Lysis Buffer (10X)980315 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).

Applications Key

ELISA: ELISA

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 G: Goat 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.

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Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

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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.
ELISA Image 1: PathScan<sup>®</sup> RP Total IRF-7 Sandwich ELISA Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.