THP1-Dual™ Cells

Notification: Reference #thpd-nfis-av can only be ordered together with reference #thpd-nfis.

NF-κB–SEAP and IRF–Lucia reporter human 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 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.

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.

THP1-Dual™ cells are resistant to the selectable markers Zeocin® and blasticidin.

Figures

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. 

Specifications

Antibiotic resistance: Zeocin®, blasticidin

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)

Guaranteed mycoplasma-free

Contents

• 3-7 x 10⁶ of THP1-Dual™ cells in a cryovial or shipping flask
• 1 ml of Normocin™ (50 mg/ml)
• 1 ml of Zeocin® (100 mg/ml)
• 1 ml of Blasticidin (10 mg/ml)
• 1 tube of QUANTI-Luc™ 4 Reagent, a Lucia luciferase detection 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, a SEAP detection reagent)

THP1-Dual™ cells are shipped on dry ice (Europe, USA, Canada, and some areas in Asia)

Description

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.

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