|THP1-Dual™ Cells||Unit size||Cat. code||Docs||Qty||Price|
NF-κB-SEAP IRF-Luc Reporter Monocytes
3-7 x 10e6 cells
NF-kB-SEAP and IRF-Lucia luciferase Reporter Monocytes
THP1-Dual™ cells were derived from the human THP-1 monocyte cell line by stable integration of two inducible reporter constructs.
As a result, THP1-Dual™ cells allow the simultaneous study of the NF-κB pathway, by monitoring the activity of SEAP, and the IRF pathway, by assessing the activity of a secreted luciferase (Lucia).
THP1-Dual™ cells induce the activation of NF-κB in response to certain TLR agonists, such as Pam3CSK4 and flagellin. They trigger the IRF pathway upon stimulation with type I IFNs and RLR or CDS agonists, such as transfected dsRNA.
Cells were pretreated or not pretreated with PMA (Phorbol 12-myristate 13-acetate; 20 ng/ml for 3 hours) and stimulated with 1 ng/ml Pam3CSK4 (TLR2), 100 ng/ml LPS-EK UP (TLR4), 100 ng/ml FLA-ST UP (TLR5), 10 µg/ml R848 (TLR7/8), 10 µg/ml Tri-DAP (NOD1), 3 μg/ml 2’3’-cGAMP (STING), 1 μg/ml poly(I:C)/LyoVec™ (RLR) or 100 ng/ml poly(dA:dT)/LyoVec™ (CDS). After 24h incubation, NF-κB and IRF activation were assessed by measuring the levels of SEAP and Lucia luciferase using QUANTI-Blue™ and QUANTI-Luc™, respectively. With QUANTI-Blue™ the levels of SEAP were determined by reading the optical density (OD) at 655 nm. With QUANTI-Luc™ the levels of Lucia luciferase were determined by measuring the relative light units (RLUs) in a luminometer.
Growth medium: RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml Normocin™, Pen-Strep (100 U/ml-100 μg/ml)
This product is covered by a Limited Use License (See Terms and Conditions).Back to the top
THP1-Dual™ is supplied with:
- 1 vial of THP1-Dual™ cells (3-7 x 106 cells) in freezing medium
- 1 ml of Normocin™ (50 mg/ml)
- 1 ml of Zeocin® (100 mg/ml)
- 1 ml of Blasticidin (10 mg/ml)
- 1 pouch of QUANTI-Luc™ (Luciferase detection medium)
- 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent)
THP1-Dual™ cells are shipped on dry ice. (Europe, USA & Canada)
See Data Sheet for storage conditions of each component.Back to the top
THP1-Dual™ cells feature the Lucia gene, a secreted luciferase reporter gene, under the control of an ISG54 minimal promoter in conjunction with five IFN-stimulated response elements.
THP1-Dual™ cells also express a secreted embryonic alkaline phosphatase (SEAP) reporter gene driven by 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.
As a result, THP1-Dual™ cells allow the simultaneous study of the NF-κB pathway, by monitoring the activity of SEAP, and the IRF pathway, by assessing the activity of Lucia luciferase.Back to the top