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HEK-Blue™ KD-TLR5 cells

HEK-Blue™ KD-TLR5 cells Unit size Cat. code Docs Qty Price
TLR5 deficient - IL1-β SEAP reporter HEK293 cells
3-7 x 10e6 cells
hkb-kdtlr5
+-
$1,142.00

TLR5 deficient, IL-1β reporter cells

HEK-Blue™ KD-TLR5 cells are designed to monitor bioactive IL-1β secreted by THP-1 cells upon flagellin-induced NLRC4 activation.

HEK-Blue™ KD-TLR5 cells are derived from the HEK293 cell line, which endogenously expresses TLR5 and the IL-1β receptor (IL-1R).

This cell line features an NF-κB-inducible SEAP reporter gene and was engineered to knock-down the expression of TLR5 to avoid activation of NF-κB upon flagellin-induced TLR5 stimulation.

The knockdown of TLR5 permits the analysis of flagellin specifically for its NLRC4 stimulating activity.

Binding of IL-1β to IL-1R initiates a signaling cascade leading to the activation of NF-κB and the subsequent production of SEAP. Detection of SEAP in the supernatant of HEK-Blue™ KD-TLR5 cells can be readily assessed using the QUANTI-Blue™ assay.


THP1-NLRC4 and HEK-Blue™ KD-TLR5 cells were co-cultured and stimulated with increasing concentrations of ultrapure flagellin from S. typhimurium (FLA-ST UP), B. subtilis (FLA-BS UP) or P. aeruginosa (FLA-PA UP), or monosodium urate (MSU, an NLRP3 inflammasome inducer which requires prior priming of THP-1 cells).

After 24h incubation, IL-1β-induced NF-κB activation was assessed by measuring the levels of SEAP using the QUANTI-Blue™ assay. When cultured alone, HEK-Blue™ KD-TLR5 cells do not respond to flagellin.

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Specifications

Antibiotic resistance: Zeocin™, puromycin

Growth Medium: DMEM, 4.5 g/l glucose, 2 mM L-Glutamine, 10% (v/v) heat-inactivated fetal bovine serum (30 min at 56ºC), 50 U/ml penicillin, 50 µg/ml streptomycin, 100 µg/ml Normocin™

Quality Control:

  • TLR5 knockdown is verified by functional assays and qRT-PCR.
  • These cells are guaranteed mycoplasma-free.
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Contents

  • 1 vial of HEK-Blue™ KD-TLR5 cells (3-7 x 106 cells) in freezing medium.
  • 100 μl Zeocin™ at 100 mg/ml.
  • 100 μl puromycin at 10 mg/ml. S
  • 1 ml Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria and fungi.
  • 1 pouch of QUANTI-Blue™ (SEAP detection medium).

Shipped on dry ice

IMPORTANT: If cells are not frozen upon arrival, contact InvivoGen immediately.

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Description

Application

HEK-Blue™ KD-TLR5 cells are designed to monitor bioactive IL-1β secreted by THP-1 cells upon flagellin-induced NLRC4 activation.

Levels of IL-1β secreted into the supernatant of THP-1 cells can be monitored using the HEK-Blue™ KD-TLR5 cell line.

THP1-NLRC4 and HEK-Blue™ KD-TLR5 cell lines can be used sequentially (option 1) or co-cultured to save time (option 2).

 

THP-1/HEK-Blue™ IL-1β Assay :

Detection of IL-1beta in THP-1 supernatants

 

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