IFN-α/β Reporter HEK 293 Cells
Product | Unit size | Cat. code | Docs. | Qty. | Price | |
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HEK-Blue™ IFN-α/β cells Human HEK293 cells - Type I IFNs Reporter Cells |
Show product |
3-7 x 10e6 cells |
hkb-ifnabv2
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HEK-Blue™ IFN-α/β Cells signaling pathway
Human Type I IFNs Reporter Cells
HEK-Blue™ IFN-α/β cells were engineered from the human embryonic kidney HEK 293 cell line to detect bioactive human type I interferons (e.g. IFN-α, IFN-β) by monitoring the activation of the JAK/ISGF3 (STAT1/STAT2/IRF9 complex) pathway.
IFN-α and IFN-β are important anti-viral cytokines that also have anti-proliferative and immunomodulatory functions [1, 2]. They bind a cell-surface receptor, composed of two subunits, IFNAR1 and IFNAR2, which are associated with TyK2 and JAK1, respectively [1].
Cell line description:
HEK-Blue™ IFN-α/β cells were generated by stable transfection with the genes encoding the human STAT2 and IRF9 to obtain a fully active type I IFN signaling pathway. The other genes of the pathway (IFNAR1, IFNAR2, JAK1, TyK2, and STAT1) are naturally expressed by these cells. HEK-Blue™ IFN-α/β cells were also stably transfected with the secreted embryonic alkaline phosphatase (SEAP) reporter under the control of the ISG54 promoter. This promoter comprises IFN-stimulated response elements (ISRE) that are recognized by the ISGF3 complex. The binding of IFN-α or IFN-β to their receptor triggers a signaling cascade leading to the activation of ISGF3 and the subsequent production of SEAP. This can be readily assessed in the supernatant using QUANTI-Blue™ Solution, a SEAP detection reagent.
HEK-Blue™ IFN-α/β cells detect human (h) IFN-α and hIFN-β. At high concentrations, they might respond to mouse (m) IFN-α, but not mIFN-β (see figures). The activation of HEK-Blue™ IFN-α/β cells can be blocked with a neutralizing monoclonal antibody, such as anti-hIFN-α-IgG. Of note, HEK-Blue™ IFN-α/β cells do not respond to human type II and type III IFNs (IFN-γ/λ) (see figures).
Key Features:
- Fully functional IFN-α/β signaling pathway
- Readily assessable SEAP reporter activity
- Strong response to human (h) IFN-α and hIFN-β
- Poor to no response to murine (m) IFN-α and mIFN-β
- Unresponsive to IFN-γ (type II IFN) and IFN-λ (type III IFN)
Applications:
- Detection of human IFN-α and IFN-β
- Screening of anti-IFN-α/β and anti-IFNAR antibodies
- Quantification of IFN-α/β activation in biological samples, such as plasma or serum [3]
References:
1. Schreiber G. 2017. The molecular basis for differential type I interferon signaling. J. Biol. Chem. 292:7285-94.
2. McNab F. et al., 2015. Type I interferons in infectious disease. Nat Rev Immunol. 15(2):87-103.
3. Gómez-Bañuelos E, et al., 2024. Uncoupling interferons and the interferon signature explain clinical and transcriptional subsets in SLE. medRxiv. 2023.08.28.23294734
Specifications
Antibiotic resistance: Blasticidin, Zeocin®
Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) heat-inactivated fetal bovine serum, 100 U/ml penicillin, 100 µg/ml streptomycin, 100 µg/ml Normocin™
Guaranteed mycoplasma-free
Detection ranges:
- 1 - 103 U/ml for human IFN-α
- 1 - 103 U/ml for human IFN-β
This product is covered by a Limited Use License (See Terms and Conditions).
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- 1 vial containing 3-7 x 106 cells
- 1 ml of Blasticidin (10 mg/ml)
- 1 ml of 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, a SEAP detection reagent)
Shipped on dry ice (Europe, USA, Canada and some areas in Asia)
Details
Type I interferons, in particular interferon-alpha (IFN-α) and interferon beta (IFN-β), play a vital role in host resistance to viral infections [1, 2]. The type I IFN family is a multi-gene cytokine family that encodes 13 partially homologous IFN-α subtypes in humans (14 in mice), a single IFN-β, and several poorly defined single gene products (IFN-ɛ, IFN-τ, IFN-κ, IFN-ω, IFN-δ, and IFN-ζ) [1, 2]. IFN-α and IFN-β are the best-defined and most broadly expressed type I IFNs [2].
IFN-β and all of the IFN-α subtypes bind to a heterodimeric transmembrane receptor composed of the subunits IFNAR1 and IFNAR2 which are associated with the tyrosine kinases Tyk2 and Jak1 (Janus kinase 1) respectively. These kinases phosphorylate STAT1 and STAT2 which then dimerize and interact with IFN regulatory factor 9 (IRF9), leading to the formation of the ISGF3 complex. ISGF3 binds to IFN-stimulated response elements (ISRE) in the promoters of IFN-stimulated genes (ISG) to regulate their expression.
Despite their protective effects, studies have shown that aberrantly expression of the type I IFN system can elicit autoimmune disorders, such as interferonopathies and SLE (systemic lupus erythematosus). Recent evidence also implicates type I IFN-dependent signaling as a key inflammatory driver in non-autoimmune diseases [3].
References:
1. Schreiber G. 2017. The molecular basis for differential type I interferon signaling. J. Biol. Chem. 292:7285-94.
2. McNab F. et al., 2015. Type I interferons in infectious disease. Nat Rev Immunol. 15(2):87-103.
3. Crow MK, Ronnblom L. Type I interferons in host defence and inflammatory diseases. Lupus Sci Med. 2019 May 28;6(1):e000336.