THP1-Blue™ NF-κB Cells
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NF-κB SEAP Reporter Monocytes
3-7 x 10e6 cells
NF-kB SEAP Reporter Monocytes
THP1-Blue™ NF-κB cells were specifically designed for monitoring the NF-κB signal transduction pathway in a physiologically relevant cell line.
THP1-Blue™ 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 determining the activity of SEAP.
The levels of NF-κB-induced SEAP in the cell culture supernatant are readily assessed with QUANTI-Blue™, a SEAP detection reagent.
THP1-Blue™ NF-κB cells are highly responsive to PRR agonists that trigger the NF-κB pathway.
Cells were incubated with 10e7 cells/ml HKLM (TLR2), 10 ng/ml Pam3CSK4 (TLR1/2), 1 ng/ml FSL-1 (TLR2/6), 10 µg/ml poly(I:C) (TLR3), 100 ng/ml LPS-EB UP (TLR4), 100 ng/ml FLA-ST UP (TLR5), 3 µg/ml CL075 (TLR8), 10 µg/ml ODN2006 (TLR9), 10 µg/ml Tri-DAP (NOD1) or 10 µg/ml MDP (NOD2). After 24h incubation, the levels of NF-κB-induced SEAP were assessed from the cell culture supernatant using QUANTI-Blue™.
Antibiotic resistance: 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)
- TLR expression was determined by RT-PCR in THP1-Blue™ NF-kB cells.
- All TLR mRNAs were detected. Considering the concentration of ligands used to stimulate these cells, TLR2, TLR1/2 and TLR2/6 responses are considered to be very strong. TLR4, TLR5 and TLR8 responses are robust.
Responses to TLR3, TLR7 and TLR9 are hardly detectable even when high concentrations of the cognate ligands are used.
- THP1-Blue™ NF-kB cells respond to NOD1 and NOD2 agonists.
- THP1-Blue™ NF-kB cells are guaranteed mycoplasma-free.
- 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 Blasticidin (10 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)
- 10e9 cells HKLM (Heat killed Listeria monocytogenes; positive control of TLR2 activity)
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THP1-Blue™ were derived from the human THP-1 monocyte cell line by stable integration of an NF-κB-inducible SEAP reporter construct.
THP1- Blue™ NF-κB cells 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-Blue™ NF-κB cells allow the monitoring of NF-κB activation by determining the activity of SEAP.