ISG Reporter HEK293 Cells - HEK-Blue™ ISG Cells

IRF-SEAP Reporter Cells

ABOUT

Interferon regulatory factor (IRF)-inducible SEAP reporter HEK293 cells

HEK-Blue™ ISG cells were specifically designed to study the activation of the STING/TBK1/IRF3 signaling pathway by cyclic dinucleotides (CDNs). 

CDNs in the cytosol bind directly to STING leading to TBK1-mediated IRF3 activation and type I interferon (IFN) production [1]. IFNs activate the JAK-STAT pathway with the subsequent activation of IFN-stimulated response elements (ISRE) in the promoters of IFN-stimulated genes (ISG).

More details More details about STING

 

Cell line description:

HEK-Blue™ ISG cells were derived from the PEAKrapid cell line (similar to ATCC® CRL-2828™) which itself was derived from the human embryonic kidney (HEK)-293 cell line.
HEK-Blue™ ISG cells express a secreted embryonic alkaline phosphatase (SEAP) under the control of the IRF-inducible promoter comprised of five IFN-stimulated response elements (ISRE) fused to an ISG54 minimal promoter.
The presence of CDNs in the cytosol of HEK-Blue™ ISG cells will induce the production of the IRF-inducible SEAP reporter directly by activating the STING/TBK1/IRF3 pathway and indirectly through the activation of the JAK/STAT/IRF9 pathway with type I IFN.

Levels of SEAP in the supernatant can be easily determined with QUANTI- Blue™ Solution, a reagent that turns purple/blue in the presence of SEAP and by reading the OD at 620-655 nm.
HEK-Blue™ ISG cells respond strongly to non-canonical CDNs, namely 2’3’-cGAMP but poorly to cytosolic DNA, DMXAA, and canonical CDNs.

Features of HEK-Blue™ ISG cells:

  • Fully functional STING/TBK1/IRF signaling pathway
  • Readily assessable SEAP reporter activity
  • Functionally tested and guaranteed mycoplasma-free

Application of HEK-Blue™ ISG cells:

  • Studying the STING/TBK1/IRF signaling pathway

 

Reference:

1. Wan D. et al., 2020. Research Advances in How the cGAS-STING Pathway Controls the Cellular Inflammatory Response. Front Immunol. 11:615.

Disclaimer:  These cells are for internal research use only and are covered by a Limited Use License (See Terms and Conditions). Additional rights may be available.

HEK-Blue™ ISG Cells signaling pathway
HEK-Blue™ ISG Cells signaling pathway

SPECIFICATIONS

Specifications

Species
Human
Tested applications

Studying the STING/TBK1/IRF signaling pathway

Cell type
Epithelial
Growth properties
Adherent
Tissue origin
Human embryonic kidney cells
Reporter gene
SEAP
Detection method
Colorimetric
Antibiotic resistance
G418 (mammalian cells)
Zeocin®
Growth medium

Complete DMEM (see TDS)

Mycoplasma-free

Verified using Plasmotest™

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    HEK-Blue™ ISG Cells
  • Cat code: 
    hkb-isg-1
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 1 ml Zeocin® (100 mg/ml)
  • 1 ml Normocin™ (50 mg/ml)
  • 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution)

Shipping & Storage

  • Shipping method:  Dry ice

    • Storage:

    • Liquid nitrogen vapor
    Stability: 20 passages

    Caution:

    • Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.

Details

STING (stimulator of interferon genes; also known as TMEM173, MITA, MPYS and ERIS) is essential for the interferon (IFN) response to cytosolic nucleic acids, such as microbial or self-DNA [1, 2], and acts as a direct sensor of cyclic dinucleotides (CDNs) [3]. CDNs are important second messenger molecules in bacteria, affecting numerous responses of the prokaryotic cell. In mammalian cells, CDNs act as agonists of the innate immune response [4]. CDNs bind directly to and activate STING leading to TANK Binding Kinase 1 (TBK1)-dependent interferon regulatory factor 3 (IRF3) activation and type I IFN production. IFNs then activate the JAK-STAT pathway with subsequent activation of IFN-stimulated response elements (ISRE) in the promoters of IFN-stimulated genes (ISG).

 

1. Wan D. et al., 2020. Research Advances in How the cGAS-STING Pathway Controls the Cellular Inflammatory Response. Front Immunol. 11:615.
2. Ishikawa H. & Barber, G., 2008. STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling. Nature 455, 674–678.
3. Burdette D.L. et al., 2011. STING is a direct innate immune sensor of cyclic di-GMP. Nature 478(7370):515-8.
4. Wu J. et al., 2013. Cyclic GMP-AMP is an endogenous second messenger in innate immune signaling by cytosolic DNA. Science 339(6121):826-30.

DOCUMENTS

Documents

HEK-Blue™ ISG Cells

Validation Data Sheet

Technical Data Sheet

Safety Data Sheet

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Certificate of analysis

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Citations

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