B16-Blue™ IFN-α/β Cells
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Cat.code:
bb-ifnt1
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ABOUT
Murine Type I IFNs Reporter Cells
B16-Blue™ IFN-α/β cells are designed to monitor mouse type I interferon (e.g. IFN-α, IFN-β)-induced JAK/ISGF3 stimulation or inhibition through SEAP activation. ISGF3 is a signaling complex comprising STAT1, STAT2, and IRF9. In addition, these cells can be used for screening antibodies or small-molecule inhibitors targeting the IFN-α/β pathway. This colorimetric cytokine bioassay can be used to screen activatory or inhibitory molecules, such as engineered cytokines and neutralizing antibodies, respectively.
Mouse IFN-α/β reporter cells respond specifically to mouse (m) IFN-α or mIFN-β (see figures). They do not respond to human (h) IFN-α/β or mIFN-γ.
Key features
- Readily assessable ISRE-inducible SEAP reporter activity
- Convenient readout using QUANTI-Blue™ Solution
- No response to human IFN-α/β or mouse IFN-γ
Applications
- Therapeutic development
- Drug screening
- Release assay
IFN-α and IFN-β are important anti-viral cytokines that also have anti-proliferative and immunomodulatory functions [1, 2].
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.
SPECIFICATIONS
Specifications
IFN-α, IFN-β
Mouse
Detection of mouse IFN-α and IFN-β
Mouse IFN-α: 102 - 104 IU/ml
Mouse IFN-β: 10 - 104 IU/ml
Complete DMEM (see TDS)
Verified using Plasmotest™
Each lot is functionally tested and validated.
CONTENTS
Contents
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Product:B16-Blue™ IFN-α/β Cells
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Cat code:bb-ifnt1
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Quantity:3-7 x 10^6 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)
Shipping & Storage
- Shipping method: Dry ice
- Liquid nitrogen vapor
- Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.
Storage:
Caution:
Details
Cell line description
B16-Blue™ IFN-α/β cells were generated from the murine B16 melanoma cell line by the stable transfection 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.
B16-Blue™ IFN-α/β cells respond specifically to murine (m) IFN-α/β and do not respond to human (h) IFN-α/β. Stimulation of these cells with mIFN-α or mIFN-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. Of note, B16-Blue™ IFN-α/β cells do not respond to mIFN-γ (see figures).
Interferon-alpha background
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.
DOCUMENTS
Documents
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
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