HEK-Blue-Lucia™ TNF-α Cells
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Cat.code:
hkd-tnfa
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ABOUT
TNF-α reporter cells with SEAP and Lucia luciferase double read-out
HEK-Blue-Lucia™ TNF-α cells (*) are designed to monitor human tumor necrosis factor-alpha (TNF-α)-induced NF-κB/AP-1 stimulation or inhibition through SEAP and Lucia® detection. It can be employed for screening both activatory molecules, such as engineered cytokines, and inhibitory molecules, such as neutralizing antibodies.
This cell line can be used as either a colorimetric or bioluminescent bioassay by monitoring the activity of SEAP and Lucia® luciferase using QUANTI-Blue™ Solution and QUANTI-Luc™ 4 Lucia/Gaussia, respectively.
HEK-Blue-Lucia™ TNF-α cells respond specifically to recombinant human TNF-α. They are not responsive to human IL-1β and IFN-β (see figures). The reliable and consistent performance of these cells makes them suitable for release assays of therapeutic molecules that inhibit TNF-α signaling, such as Adalimumab, a monoclonal antibody targeting TNF-α.
Key features
- Fully functional TNF-α signaling pathway
- Readily assessable NF-κB-inducible SEAP and Lucia luciferase reporter activities
- Strong response to humanTNF-α
- No response to human IL-1β or hIFN-β
Applications
- Therapeutic development
- Drug screening
- Release assay
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].
(*) Note: This cell line has been renamed. It was formerly known as "HEK-Dual™ TNF-α". The cat. code (hkd-tnfa) remains unchanged.
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
TNF-α
Detection and quantification of human TNF-α activity
Using QUANTI-Blue™ Solution: 1 ng - 1 µg/ml
Using QUANTI-Luc™ 4 Gaussia/Lucia®: 0.5 ng - 1 µg/ml
Complete DMEM (see TDS)
Verified using Plasmotest™
Each lot is functionally tested and validated.
CONTENTS
Contents
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Product:HEK-Blue-Lucia™ TNF-α Cells
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Cat code:hkd-tnfa
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Quantity:3-7 x 10^6 cells
- 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)
Shipping & Storage
- Shipping method: Dry ice
- Liquid nitrogen vapor
- Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.
Storage:
Caution:
Details
Cell line description
HEK-Blue-Lucia™ TNF-α cells were generated from human embryonic kidney cells (HEK293) through the 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 activation 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.
TNF-α Background
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].
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.
DOCUMENTS
Documents
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
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