B16-Blue™ IFN-α/β Cells

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Murine Type I IFNs Sensor Cells

B16-Blue™ IFN-α/β cells signaling

B16-Blue™ IFN-α/β cells allow the detection of bioactive murine type I IFNs  (i.e. IFN-α and IFN-β) by monitoring the activation of the JAK/STAT/ISGF3 pathway and/or IRF3 pathway.

IFN-α and IFN-β are important anti-viral cytokines that also have anti-proliferative and immunomodulatory functions [1, 2]. These cytokines bind a cell-surface receptor, composed of two subunits, IFNAR1 and IFNAR2, which are associated with TyK2 and JAK1, respectively [1]. Upon binding to this receptor, type I IFNs trigger the JAK/STAT/ISGF3 pathway.

More details

Cell line description:

B16-Blue™ IFN-α/β cells derive from the murine B16 melanoma cell line of C57BL/6 origin after stable transfection with a SEAP (secreted embryonic alkaline phosphatase) reporter gene under the control of the IFN-α/β-inducible ISG54 promoter enhanced by a multimeric ISRE. These cells do not respond to IFN-γ, due to the inactivation of the IFN-γ receptor.

B16-Blue™ IFN-α/β cells respond specifically to mIFN-α/β and do not respond to human IFN-α/β. Stimulation of these cells with murine IFN-α or IFN-β, or type I IFN inducers, such as poly(I:C), poly(dA:dT) or 5’ppp-dsRNA delivered intracellularly, triggers the production of SEAP  by the activation of the IRF-inducible promoter. Levels of SEAP can be easily monitored using the detection medium QUANTI-Blue™ Solution.

Features of B16-Blue™ IFN-α/β cells:

• Fully functional murine IFN-α/β signaling pathway
• Do not respond to human IFN-α/β
• Do not respond to murine IFN-γ (type II IFN)
• Readily assessable SEAP reporter activity
• Functionally tested and guaranteed mycoplasma-free

Applications of B16-Blue™ IFN-α/β cells:

• Detection of murine IFN-α and IFN-β 
• Screening of anti-mIFN-α or anti-mIFN-β antibodies

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.

Figures

Dose-response of B16-Blue™ IFN‑α/β cells to recombinant murine IFN‑α/β.
Cells were stimulated with increasing concentrations of recombinant murine IFN‑αA (also known as mIFN-α3) and mIFN-β. After overnight incubation, the ISGF3 response was determined using QUANTI‑Blue™ Solution, a SEAP detection reagent, and reading the optical density (OD) at 630 nm. The OD at 630 nm is shown as mean ± SEM.

Response of B16-Blue™  IFN-α/β cells to a panel of cytokines.
Cells were stimulated with various human and murine recombinant cytokines: 1000 IU/ml mIFN-αA, mIFN-β, human IFN-α2a (hIFN-α2a), hIFN-β, 100 ng/ml of mIFN-λ, mIFN-γ, hIFN-γ, or 1 µg/ml of the type I IFN inducer poly(dA:dT) complexed extemporaneously with the transfection reagent LyoVec™ (Poly(dA:dT/LV). After overnight incubation, SEAP activity was assessed using QUANTI-Blue™ Solution. The OD at 630 nm is shown as mean ± SEM.

Specifications

Detects murine type I interferons:

• Detection range for mouse IFN-α: 10² - 10⁴ IU/ml
• Detection range for mouse IFN-β: 10² - 10⁴ IU/ml

Antibiotic resistance: Zeocin®

Growth medium: DMEM, 10% (v/v) heat-inactivated fetal bovine serum, 2 mM L-glutamine, 100 µg/ml Normocin™, 100 U/ml penicillin, 100 µg/ml streptomycin

Guaranteed mycoplasma-free

This product is covered by a Limited Use License (See Terms and Conditions).

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml of Zeocin® (100 mg/ml)
• 1 ml of 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 14 partially homologous IFN-α subtypes in mice (13 in humans), 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. 

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.

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