THP1-Blue™ NF-κB Cells

Notification: Reference #thp-nfkbv2-av can only be ordered together with reference #thp-nfkbv2.

NF-κB–SEAP Reporter Monocytes

Signaling pathways in THP1-Blue™ NF-κB cells
(click to enlarge)

THP1-Blue™ NF-κB cells were specifically designed for monitoring the NF-κB signal transduction pathway in a physiologically relevant cell line. THP1-Blue™ cells were derived from the human THP-1 monocyte cell line by stable integration of an NF-κB-inducible SEAP reporter construct.

As a result, THP1-Blue™ NF-κB cells allow the monitoring of NF-κB activation by assessing the activity of SEAP (secreted embryonic alkaline phosphatase). The level of NF-κB-induced SEAP in the cell culture supernatant is readily assessed with QUANTI-Blue™, a SEAP detection reagent.

THP-1 cells endogenously express many pattern-recognition receptors (PRRs), including Toll-like receptors (TLRs), the stimulator of interferon genes (STING), and RIG-I-like receptors (RLR). Therefore, THP1-Blue™ NF-κB cells are highly responsive to PRR agonists that trigger the NF-κB pathway (see figure).

Key features

• Functionally validated with a selection of PRR ligands and cytokines
• Readily assessable NF-κB activation by assessing the SEAP activity

Note: A new clone is provided with an improved reporter/background signal ratio. The cat code has been changed accordingly (thp-nfkbv2).

Figures

NF-κB responses in THP1-Blue™ NF-κB cells. Cells were incubated for 24 hours with various PRR ligands: Pam3CSK4 (TLR2 ligand, 1 ng/ml), Poly(I:C) HMW (TLR3 ligand, 10 µg/ml), LPS-EK Ultrapure (UP) (TLR4 ligand, 1 ng/ml), FLA-ST UP (TLR5 ligand, 1 ng/ml), Imiquimod (TLR7 ligand, 10 µg/ml), R848 (TLR7/8 ligand, 10 µg/ml), TL8-506 (TLR8 ligand, 1 µg/ml), ODN 2006 (TLR9 ligand, 10 µg/ml), Tri-DAP (NOD1 ligand, 10 µg/ml), MDP (NOD2 ligand, 10 µg/ml), 2’3’-cGAMP (STING ligand, 10 µg/ml), and 3p-hpRNA complexed with LyoVec™ (LV) (RIG-I ligand, 1 µg/ml). Human TNF-α (1 ng/ml) was used as an NF-κB-positive control. After 24h incubation, the NF-κB-induced SEAP activity was assessed. Data are shown as OD at 650 nm (mean ± SEM).

Specifications

Antibiotic resistance: Zeocin® 

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)

Quality Control:

• Reporter activity has been verified by functional assays.
• The stability for 20 passages, following thawing, has been verified. 
• These cells are guaranteed mycoplasma-free. 

Please note, that the old clone purchased before March 2024 (cat. code thp-nfkb) is selectable with Blasticidin. For more details, please read the technical data sheet.

Contents

• 1 vial of THP1-Blue™ NF-κB 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 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 and some areas in Asia)

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