J774-Dual™ Cells
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Cat.code:
j774d-nfis
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ABOUT
NF-kB-SEAP & IRF-Lucia Reporter Mouse Macrophages
J774-Dual™ cells have been derived from the mouse J774.1 macrophage-like cell line by stable integration of two inducible reporter constructs.
J774-Dual™ cells express a secreted embryonic alkaline phosphatase (SEAP) reporter gene under the control of an IFN-β minimal promoter fused to five copies of the NF-κB consensus transcriptional response element and three copies of the c-Rel binding site.
J774-Dual™ cells also express the Lucia®luciferase gene, which encodes a secreted luciferase, under the control of an ISG54 minimal promoter in conjunction with five IFN-stimulated response elements.
As a result, J774-Dual™ cells allow to simultaneously study the NF-κB pathway, by assessing the activity of SEAP, and the interferon regulatory factor (IRF) pathway, by monitoring the activity of Lucia luciferase.
Both reporter proteins are readily measurable in the cell culture supernatant when using QUANTI-Blue™ Solution, a SEAP detection reagent, and QUANTI-Luc™ 4 Lucia/Gaussia, a Lucia and Gaussia luciferase detection reagent.
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
Detection and quantification of the NF-κB and IRF pathway activation
Complete DMEM (see TDS)
Verified using Plasmotest™
Each lot is functionally tested and validated.
CONTENTS
Contents
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Product:J774-Dual™ Cells
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Cat code:j774d-nfis
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Quantity:3-7 x 10^6 cells
1ml of Blasticidin (10 mg/ml)
1 ml of Zeocin® (100 mg/ml)
1 ml of& Normocin™ (50 mg/ml)
1 tube of QUANTI-Luc™ 4 Reagent (sufficient to prepare 25 ml)
1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution)
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
J774.1 cells express a variety of pattern recognition receptors (PRRs), including Toll-like receptors (TLRs) [1, 2], C-type lectin receptors (CLRs) [2,3], RIG-I-like receptors (RLRs) [4].
Upon recognition of their cognate PAMPs, these receptors induce signaling pathways leading to the activation of the
transcription factors NF-kB and/or IRF3/7. Stimulation of J774-Dual™ cells with the following PAMPs, Pam3CSK4 (TLR1/2), lipopolisaccharide (TLR4), CpG ODNs (TLR9), L18-MDP (NOD2) and TDB (Mincle), leads to the activation of NF-kB.
Stimulation with RLR ligands, such as transfected poly(I:C) or 5’ppp-dsRNA, or the STING agonist, 2’3’-cGAMP, triggers the IRF pathway.
J774-Dual™ cells are resistant to the selectable markers blasticidin and Zeocin®.
1. Jin M. et al., 2011. Effects of chondroitin sulfate and its oligosaccharides on toll-like receptor-mediated IL-6 secretion by macrophage-like J774.1 cells. Biosci Biotechnol Biochem. 75(7):1283-9.
2. Kushida T. et al., 2011. Enhancement of Dectin-2 gene expression by lignin-carbohydrate complex from Lentinus edodes mycelia extract (LEM) in a mouse macrophage-like cell line. Anticancer Res. 31(4):1241-8.
3. Takeda Y. et al., 2007. Ternary complex consisting of DNA, polycation, and a natural polysaccharide of schizophyllan to induce cellular uptake by antigen presenting cells. Biomacromolecules. 8(4):1178-86.
4. Wilden H. et al., 2009. Expression of RIG-I, IRF3, IFN-beta and IRF7 determines resistance or susceptibility of cells to infection by Newcastle Disease Virus. Int J Oncol. 34(4):971-82.
DOCUMENTS
Documents
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
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