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RAW-Blue™ ISG Cells

RAW-Blue™ ISG cells Unit size Cat. code Docs Qty Price
Mouse Macrophage IFN Reporter Cells
3-7 x 10e6 cells
raw-isg
+-
$1,071.00

April 17th 2019 - CONTENTS UPDATE NOTIFICATION
Please note that, for your convenience, these cells are now provided with QUANTI-Blue™ Solution.

Mouse Macrophage IFN Reporter Cells

RAW-Blue™ ISG cells were designed for the detection of bioactive murine type I IFNs in a physiologically relevant cell line.

RAW-Blue™ ISG cells were derived from the murine RAW 264.7 macrophage cell line by stable integration of an interferon regulatory factor (IRF)-inducible secreted embryonic alkaline phosphatase (SEAP) reporter construct.

The levels of IRF-induced SEAP in the cell culture supernatant are readily assessed with QUANTI-Blue™, a SEAP detection reagent. 

RAW-Blue™ ISG cells are responsive to murine IFN-α and IFN-β but do not respond to their human counterparts.

They are also responsive to PRR ligands that trigger the IFN signaling pathway, such as transfected double-stranded nucleic acids.

Figures


Responses of RAW-Blue™ ISG cells to type I IFNs and to transfected double-stranded nucleic acid
RAW-Blue™ cells were incubated with increasing concentrations of mouse or human recombinant IFN-α or IFN-β, poly(dA:dT)/LyoVec or poly(dG:dC)/LyoVec.
After 24h incubation the levels of IRF-induced SEAP were determined using QUANTI-Blue™.

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Specifications

  • Detection range for mIFN-α: 5x10e2 - 5x10e4 IU/ml
  • Detection range for mIFN-β: 10e1 - 10e3 IU/ml

Antibiotic resistance: Zeocin™

Growth medium: DMEM medium, 2 mM L-glutamine, 10% FBS supplemented with 100 µg/ml Normocin™ and 200 µg/ml Zeocin™.

Quality control:

  • Activity of RAW-Blue™ ISG cells was tested with murine interferon alpha (mIFN-α), murine interferon beta (mIFN-β) and pathogen-associated molecular patterns (PAMPs), such as Poly(I:C).
  • RAW-Blue™ ISG cells are guaranteed mycoplasma-free.
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Contents

  • 1 vial containing 3-7 x 106 cells
  • 100 μl 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)

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Description

RAW-Blue ISG pathway

RAW-Blue™ ISG cells allow the detection of bioactive murine type I IFNs or PAMPs by monitoring the activation of the interferon regulatory factor (IRF) pathway.

These cells are derived from the RAW 264.7 macrophages. RAW-Blue™ ISG cells stably express a secreted embryonic alkaline phosphatase (SEAP) gene under the control of the interferon-stimulated gene 54 (ISG54)-inducible promoter enhanced by a multimeric IFN-stimulated response elements (ISRE).

Stimulation of RAW-Blue™ ISG cells with mIFN-α or mIFN-β, or type I IFN inducers, such as tranfected poly(dA:dT) or 5’ppp-dsRNA, can activate the IRF3, 7 or 9 pathways and trigger the subsequent production of SEAP.

Levels of SEAP in the supernatant can be easily determined with QUANTI-Blue™, a medium that turns purple/blue in the presence of SEAP and by reading the OD at 655 nm.

RAW-Blue™ ISG cells are resistant to Zeocin™.

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Citations

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