Invivogen
Menu

HEK-Blue-Lucia™ TNF-α Cells

Product Unit size Cat. code Docs. Qty. Price

HEK-Blue-Lucia™ TNF-α Cells

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

Show product

3-7 x 10e6 cells

hkd-tnfa
+-
$1,457

HEK-Blue-Lucia™ TNF-α vial

Additional cell vial

Show product

3-7 x 10e6 cells

hkd-tnfa-av
+-

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]. 

more More details

* 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

Applications:

  • 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

 

 

Reference:

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

Figures

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).

Back to the top

Specifications

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).

Back to the top

Contents

  • 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)

Back to the top

Details

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.

 

Back to the top

FAQ Cell Lines

Visit our FAQ Any questions about our cell lines ? Visit our frequently asked questions page

Back to the top

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.

Customer Service
& Technical Support
Shopping cart is empty