Human TLR2-KO Dual Reporter THP-1 Cells

NF-κB-SEAP and IRF-Lucia reporter monocytes

ABOUT

TLR2 knockout dual reporter monocytes

InvivoGen offers a human monocyte-derived cell line specifically designed for the study of human TLR2 (Toll-like receptor 2) function:

• THP1-Dual™ KO-TLR2 cells

THP1-Dual™ KO-TLR2 cells were generated from the THP1-Dual™ cell line through the stable knockout of the TLR2 gene. They feature two inducible reporter genes, allowing the concomitant study of the IRF and NF-κB pathways, by monitoring the Lucia luciferase and SEAP (secreted embryonic alkaline phosphatase) activities, respectively.
As expected, the NF-κB-mediated response is abolished in THP1-Dual™ KO-TLR2 cells upon incubation with TLR2-specific ligands such as Pam3CSK4 (TLR2/1), FSL-1 (TLR2/6), and heat-killed Listeria monocytogenes (HKLM; TLR2/6) when compared to the THP1-Dual™ cells, with no notable difference for the other ligands tested. Additionally, as TLR2 does not directly signal through an IRF-dependent pathway, the secretion of Lucia luciferase is unaltered in THP1-Dual™ KO-TLR2 when tested across a range of IRF-inducing ligands (see Figures).

 

Background:

Toll-like receptor 2 (TLR2) plays an essential role in detecting a diverse range of microbial pathogen-associated molecular patterns (PAMPs) from bacteria, fungi, and parasites, including lipoproteins, lipoteichoic acid, lipoarabinomannan, and chitin [1]. A number of viruses have also been shown to interact directly with TLR2, including HIV and herpes simplex virus [1, 2]. TLR2 forms a heterodimer on the cell surface with either of its co-receptors, TLR1 or TLR6, which is crucial for signaling and ligand specificity. The TLR2/TLR1 and TLR2/TLR6 heterodimers specifically bind lipoproteins depending on whether they are tri- or diacylated, respectively [1]. Their activation triggers NF-κB- and AP-1-mediated pro-inflammatory responses [3].

More Learn more about TLR2 heterodimers.

Key Features:

  • Verified knockout of the TLR2 gene (PCR, DNA sequencing, and functional assays)
  • Functionally validated with a selection of PRR ligands and cytokines
  • Readily assessable Lucia luciferase and SEAP reporter activities
  • Stability guaranteed for 20 passages
     

Applications:

  • Defining the role of TLR2 in PRR-induced signaling, or related cell signaling pathways
  • Exclusion of contaminating TLR2 agonist-dependent (e.g. bacterial lipoproteins) signaling
  • Highlighting possible overlap between TLR2 and other signaling pathways

 

References

1. Oliveira-Nascimento, L. et al. 2012. The Role of TLR2 in Infection and Immunity. Front Immunol 3, 79.
2. Henrick, B.M. et al. 2015. HIV-1 Structural Proteins Serve as PAMPs for TLR2 Heterodimers Significantly Increasing Infection and Innate Immune Activation. Front Immunol 6, 426.
3. Li, J. et al. 2013. Evolving Bacterial Envelopes and Plasticity of TLR2-Dependent Responses: Basic Research and Translational Opportunities. Front Immunol 4, 347.

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

Species
Human
Tested applications

Screening of PRR agonists or inhibitors

Cell type
Monocytic
Growth properties
Suspension
Tissue origin
Human monocytes
Reporter gene
SEAP
Lucia®
Detection method
Colometric (SEAP), Bioluminescence (Lucia)
Antibiotic resistance
Blasticidin
Zeocin®
Growth medium

Complete RPMI 1640 (see TDS)

Mycoplasma-free

Verified using Plasmotest™

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    THP1-Dual™ KO-TLR2 Cells
  • Cat code: 
    thpd-kotlr2
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria, and fungi.
  • 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)

Shipping & Storage

  • Shipping method:  Dry ice
  • Storage:

    • Liquid nitrogen vapor
    Stability: 20 passages

DOCUMENTS

Documents

THP1-Dual™ KO-TLR2 Cells

Technical Data Sheet

Validation Data Sheet

Safety Data Sheet

Certificate of analysis

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