HEK-Blue™ IFN-α/β Cells
|HEK-Blue™ IFN-α/β cells||Unit size||Cat. code||Docs||Qty||Price|
Human HEK293 cells - Type I IFNs reporter cells
3-7 x 10e6 cells
Human Type I IFN Sensor Cells
HEK-Blue™ IFN-α/β cells allow the detection of bioactive human type I IFNs by monitoring the activation of the ISGF3 pathway.
These cells were generated by stable transfection of HEK293 cells with the human STAT2 and IRF9 genes 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 in sufficient amounts.
The cells were further transfected with a SEAP reporter gene under the control of the IFN-α/β inducible ISG54 promoter. Stimulation of HEK-Blue™ IFN-α/β cells with human IFN-α or IFN-β activates the JAK/STAT/ISGF3 pathway and subsequently induces the production of SEAP.
Levels of SEAP in the supernatant can be easily determined with QUANTI-Blue™.
Stimulation of HEK-Blue™ IFN-α/β cells with human IFN-α can be blocked by anti-hIFN-α-IgA, a neutralizing monoclonal antibody.
Stimulation of HEK-Blue™ IFN-α/β cells by recombinant human IFN-α, IFN-β and IFN-γ. After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ and reading the optical density (O.D.) at 655 nm.
|hIFN-α||75 +/- 10 IU/ml||20|
|hIFN-β||10 +/- 2 IU/ml||20|
Note: The response ratio was calculated by dividing the OD at 655 nm for the treated cells by the OD at 655 nm for the untreated cells.
HEK-Blue™ IFN-α/β cells were stimulated with various human recombinant cytokines; IFNα (1000 IU/ml), IFNβ (1000 IU/ml), IFNγ (1000 IU/ml), IL-1β (100 ng/ml), IL-4 (100 ng/ml), IL-6 (100 ng/ml), IL-13 (100 ng/ml), IL-18 (100 ng/ml), TGF-β (10 ng/ml), and TNF-α (100 ng/ml). After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ and reading the O.D. at 655 nm.
Anti-hIFN-α-IgA was incubated with 500 IU/ml of human IFN-α for 30 minutes prior to the addition of HEK-Blue™ IFN-α/β cells, which were incubated with the antibody and cytokine for a further 24 h. Levels of SEAP in the supernatant were measured using QUANTI-Blue™ and reading the O.D. at 655 nm.
Growth medium: DMEM, 4.5 g/l glucose, 2-4 mM L-glutamine, 10% (v/v) fetal bovine serum, 50 U/ml penicillin, 50 μg/ml streptomycin, 100 μg/ml Normocin™
Detects human type I interferons:
- Detection range for human IFN-α: 1 - 104 IU/ml
- Detection range for human IFN-β: 10 - 104 IU/ml
- 1 vial containing 3-7 x 106 cells
- 100 μl Blasticidin (10 mg/ml)
- 100 μl Zeocin™ (100 mg/ml)
- 1 ml Normocin™ (50 mg/ml)
- 1 pouch of QUANTI-Blue™ (SEAP detection medium)
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Type I interferons, in particular interferon alpha (IFN-α) and interferon beta (IFN-β), play a vital role in host resistance to viral infections.
They signal mainly through the JAK-STAT pathway. Following their production, IFN-α and IFN-β bind to a common receptor (IFNAR) and recruit the Janus kinases (JAK1 and TyK2).
JAKs phosphorylate STAT1 and STAT2, which then dimerize and interact with IFN regulatory factor 9 (IRF9), forming a complex named ISGF3.
ISGF3 binds to IFN-stimulated response elements (ISRE) in the promoters of IFN-stimulated genes (ISG) to regulate their expression.
Stimulation of HEK-Blue™ IFN-α/β cells with human IFN-α or IFN-β activates the JAK/STAT/ISGF3 pathway and subsequently induces the production of SEAP.Back to the top
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