RAW-Dual™ Cells

RAW-Dual(IRF-Lucia/KIN-[MIP-2]SEAP) cells are derived from RAW 264.7 macrophages, a murine immune cell model that expresses many PRRs such as the Toll-like receptors (TLRs) TLR2 [1] and TLR4 [2], and the cyclic dinucleotide sensor STING [3]. RAW-Dual™ cells respond to PRR ligands that trigger the NF-kB and the IRF pathways. They also respond to murine, but not human, IFN-α and IFN-β. These cells stably express two different secreted reporter genes: the SEAP (secreted embryonic alkaline phosphatase) gene and the Lucia luciferase gene, which encodes a novel secreted luciferase.

The Lucia luciferase gene is under the control of an ISG54 minimal promoter in conjunction with five IFN-stimulated response elements. The SEAP gene expression depends on the activation of the MIP-2 promoter. MIP-2, the murine ortholog of IL-8, is a chemokine which is produced in an NF-kB dependent-manner [4]. The MIP-2 ORF has been replaced by the SEAP ORF, while the MIP-2 promoter has been left intact; thus, these cells produce SEAP upon activation of the endogenous MIP-2 promoter but do not produce MIP-2. Both reporter proteins are readily measurable in the cell culture supernatant by using QUANTI-Blue™, a SEAP detection reagent, and QUANTI-Luc™, a Lucia luciferase detection reagent. As a result, RAW-Dual™ cells allow to simultaneously study the NF-kB pathway, by assessing the activity of SEAP, and the IRF pathway, by monitoring the activity of Lucia luciferase.

RAW-DualCells are resistant to  Zeocin™.

Figures for this product

IRF INDUCTION (Lucia luciferase reporter)MIP-2 (NF-kB) INDUCTION (SEAP reporter)


The biallelic replacement of the mouse MIP-2 (also known as CXCL2) gene with the SEAP reporter gene was verified by PCR and sequencing. Furthermore, the inability to produce MIP-2 has been confirmed by ELISA.

Activity of RAW-Dualcells was tested with pattern recognition receptor (PRR) ligands that trigger the NF-kB and interferon regulatory factor (IRF) signaling pathways.

The stability of this cell line for 20 passages following thawing has been verified.

Antibiotic resistance: Zeocin™

Guaranteed mycoplasma-free

Growth medium: DMEM, 2 mM L-glutamine, 4.5 g/l glucose, 10% FBS supplemented with 100 µg/ml Normocin™ .

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


1 vial containing 3-7 x 106 cells

100 μl Zeocin™ (100 mg/ml)

1 ml Normocin™ (50 mg/ml)

1 pouch of QUANTI-Blue™ (100 ml)

1 pouch of QUANTI-Luc™ (100 ml)

Shipped on dry ice


RAW-Dual pathway


  1. Underhill DM. et al., 1999. The Toll-like receptor 2 is recruited to macrophage phagosomes and discriminates between pathogens. Nature. 401:811-5.
  2. Hornef MW. et al., 2003. Intracellular Recognition of Lipopolysaccharide by Toll-like Receptor 4 in Intestinal Epithelial Cells. J Exp Med.198:1225-35..
  3. Tanaka Y. & Chen ZJ., 2012. STING specifies IRF3 phosphorylation by TBK1 in the cytosolic DNA signaling pathway. Sci Signal. 5(214):ra20.
  4. Kim DS. et al., 2003. NF-kappaB and c-Jundependent regulation of macrophage inflammatory protein-2 gene expression in response to lipopolysaccharide in RAW 264.7 cells. Mol Immunol. 40(9):633-43.


RAW-Dual™ Cells

Description Dual IRF and MIP-2 (NF-kB) reporter mouse macrophages
Cat. Coderawd-ismip
Unit Size3-7 x 10e6 cells
Price For price or distributor address,
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