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

Tested applications

Studying the STING/TBK1/IRF signaling pathway

Species
Human
Cell type
Epithelial
Growth properties
Adherent
Tissue origin
Human embryonic kidney cells
Reporter gene
SEAP
Detection method
Colorimetric
Growth medium

Complete DMEM (see TDS)

Antibiotic resistance
G418 (mammalian cells)
Zeocin®
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

Technical Data Sheet

Validation Data Sheet

Safety Data Sheet

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Certificate of analysis

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