HEK-Blue-Lucia™ TNF-α Cells

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HEK-Blue-Lucia™ TNF-α Cells

Human TNF-α double NF-κB–readout reporter cells

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3-7 x 10e6 cells


HEK-Blue-Lucia™ TNF-α vial

Additional cell vial

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3-7 x 10e6 cells


Notification:  Reference #hkd-tnfa-av can only be ordered together with reference #hkd-tnfa.

TNF-α reporter cells with SEAP and Lucia luciferase double read-out

Signaling pathways in HEK-Blue-Lucia™ TNF-α cells
Signaling pathways in HEK-Blue-Lucia™ TNF-α cells

HEK-Blue-Lucia™ TNF-α* cells enable the detection of bioactive human tumor necrosis factor-alpha (TNF-α) by monitoring the activation of the NF-κB pathway. TNF-α is a multi-functional pro-inflammatory cytokine involved in the regulation of a wide spectrum of biological processes, such as cell proliferation, differentiation, and apoptosis [1]. 

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* formerly named HEK-Dual™ TNF-α cells


Cell line description:

HEK-Blue-Lucia™ TNF-α cells were generated by stable transfection with the genes encoding for two NF-κB-inducible reporters: secreted embryonic alkaline phosphatase (SEAP) and Lucia luciferase. This feature allows the double readout of the NF-κB actiavtion upon the binding of TNF-α to its receptor, by monitoring the activity of SEAP and Lucia luciferase using QUANTI-Blue™ Solution and QUANTI-Luc™ 4 Lucia/Gaussia, respectively. 

HEK-Blue™ TNF-α cells detect human TNF-α. Of note, these cells are not responsive to hIL-1β (see figures).

Key features:

  • Fully functional TNF-α signaling pathway
  • Readily assessable NF-κB-inducible SEAP and Lucia luciferase reporter activities
  • Do not respond to IL-1β


  • Detection and quantification of human TNF-α activity
  • Screening of anti-TNF-α or anti-TNFR1/TNFR2 antibodies
  • Screening of small molecule inhibitors of the TNF-α pathway




1.Steeland S, Libert C, Vandenbroucke RE. 2018. A New Venue of TNF Targeting. Int J Mol Sci.;19(5):1442. 


Cellular response to human TNF-α
Cellular response to human TNF-α

Dose-response of HEK-Blue-Lucia™ TNF-α cells to recombinant human TNF-α. Cells were stimulated overnight with increasing concentrations of recombinant hTNF-α. The NF-κB-induced SEAP (A) and Lucia (B) activities were assessed using QUANTI-Blue™ and QUANTI-Luc™ 4 Lucia/Gaussia detection reagents, respetively. Data are shown as optical density (OD) at 630 nm, or fold response (mean ± SEM).

HEK-Blue-Lucia™ TNF-α specificity
HEK-Blue-Lucia™ TNF-α specificity

Response profile of HEK-Blue-Lucia™ TNF-α cells. Cells were incubated overnight with recombinant hTNF-α (10 ng/ml), hIFN-β (1000 U/ml), or hIL-1β (1 µg/ml). The NF-κB-induced SEAP (A) and Lucia (B) activities were assessed using QUANTI-Blue™ and QUANTI-Luc™ 4 Lucia/Gaussia detection reagents, respetively. Data are shown as optical density (OD) at 630 nm, or fold response (mean ± SEM).

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Antibiotic resistance: Zeocin®

Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) heat-inactivated fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 100 μg/ml Normocin™

Quality Control:

  • Reporter gene activities in response to TNF-α and various cytokines have been validated using functional assays.
  • The stability for 20 passages, following thawing, has been verified.
  • These cells are guaranteed mycoplasma-free. 

Detection range for hTNF-α when using:

  • QUANTI-Blue™ Solution: 1 ng - 1 µg/ml
  • QUANTI-Luc™ 4 Gaussia/Lucia: 0.5 ng - 1 µg/ml


These cells are covered by a Limited Use License (See Terms and Conditions).

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  • 3-7 x 106 cells in a cryovial or shipping flask.
  • 1 ml of Zeocin® (100 mg/ml).
  • 1 ml of Normocin™ (50 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)
  • 1 tube of QUANTI-Luc™ 4 Reagent, a Lucia luciferase detection reagent (sufficient to prepare 25 ml)

Dry ice Shipped on dry ice (Europe, USA, Canada, and some areas in Asia)

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Tumor necrosis factor-alpha (TNF-α) is a pleiotropic inflammatory cytokine produced by several types of cells, predominantly activated macrophages [1.] TNF-α plays an important role in the immune response to microbial invasions and in the necrosis of specific tumors. Of note,  as a potent mediator of inflammation, TNF-α has been implicated in the pathogenesis of several autoimmune and inflammatory diseases such as rheumatoid arthritis and inflammatory bowel disease [1,2].

TNF-α exists in two forms; a type II transmembrane protein and a mature soluble protein. The TNF-α transmembrane protein is proteolytically cleaved to yield a soluble protein [3], which subsequently forms a non-covalently linked homotrimer in solution. TNF-α binds two receptors TNFR1 and TNFR2 inducing signaling that involves TRADD, TRAF2, and RIP, and leads to the activation of the NF-κB and the MAPK pathways [4]. 

Interleukin 1 beta (IL-1β) is another inflammatory cytokine that triggers these pathways following the binding to its receptor IL-1RI and the recruitment of MyD88. Both TNF-α and IL-1β receptors are expressed in HEK293 cells. HEK-Dual™ TNF-α Cells are rendered unresponsive to IL-1β by stable knock-out of the MyD88 gene.


1. Sedger L. & McDermott M., 2014. TNF and TNF-receptors: From mediators of cell death and inflammation to therapeutic giants - past, present and future. Cytokine Growth Factor Rev. 25(4):453-72.
2. Li P. et al., 2017. Drugs for Autoimmune Inflammatory Diseases: From Small Molecule Compounds to Anti-TNF Biologics.Front Pharmacol .8:460.
3. Kriegler M. et al., 1988. A novel form of TNF/cachectin is a cell surface cytotoxic transmembrane protein: ramifications for the complex physiology of TNF. Cell. 53(1):45-53.
4. Wajant H. et al., 2003. Tumor necrosis factor signaling. Cell Death Differ. 2003 10(1):45-65.


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Notification: This cell line has been renamed. It was formerly known as "HEK-Dual™ TNF-α". The cat. code (hkd-tnfa) remains unchanged.
This product is for internal research use only. Additional rights may be available. Please visit InvivoGen’s Terms and Conditions.

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