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B16-Blue™ ISG Cells

B16-Blue™ ISG Cells Unit size Cat. code Docs Qty Price
Murine B16 melanoma - IFNs reporter cells
3-7 x 10e6 cells
bb-ifnabg
+-
$1,035.00

Interferon Regulatory Factor-Inducible SEAP Reporter B16 Melanocytes

B16-Blue™ ISG cells allow the detection of bioactive murine type I and II IFNs by monitoring the activation of the JAK/STAT pathway.
B16-Blue™ ISG cells can also be used study the activation of the TBK1/IRF3 pathway by cytosolic DNA, dsRNA or CDNs.

B16-Blue™ ISG cells were derived from the murine B16 F1 melanoma cell line. They express the secreted embryonic alkaline phosphatase (SEAP) reporter gene under the control of the I-ISG54 promoter which is comprised of the IFN-inducible ISG54 promoter enhanced by a multimeric ISRE.
Stimulation of B16- Blue™ ISG cells with IFNs, CDNs, such as cGAMP, or type I IFN inducers, such as transfected poly(dA:dT), triggers the activation of the I-ISG54 promoter and the production of SEAP.
Levels of SEAP in the supernatant can be easily determined using QUANTI- Blue™, a reagent that turns purple/blue in the presence of SEAP, by reading the OD at 620-655 nm.
B16-Blue™ ISG cells are resistant to Zeocin™.


Response of B16-Blue ISG and B16-Blue ISG-KO-STING to CDNs and IFN-β:
Cells were stimulated with 30 µg/ml of the cyclic dinucleotides, and 103 
U/ml of mIFN-β. Cells were not permeabilized.
After 24h incubation, the levels of IRF-induced SEAP or Lucia luciferase were determined using QUANTI-Blue™.

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Specifications

Murine IFN Detection range :
• mIFN-α: 10e2 - 10e4 IU/ml
• mIFN-β: 10e2 - 10e4 IU/ml
• mIFN-γ: 0.1 ng - 1 µg/ml

Antibiotic resistance: Zeocin™

Growth Medium: RPMI, 10% (v/v) fetal bovine serum (FBS), 50 U/ml penicillin, 50 µg/ml streptomycin, 100 µg/ml Normocin™, 2 mM L-glutamine

Quality control:

  • Reporter activity is validated by stimulating the cells with murine IFN-α (mIFN-α), mIFN-β, mIFN-γ, and IRF3 activators, such as poly(dA:dT)/LyoVec™, c-di-GMP and cGAMP.
  • The cells are guaranteed mycoplasma-free.
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Contents

Shipped on dry ice

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