HEK-Blue™ TNF-α Cells
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Cat.code:
hkb-tnfdmyd
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ABOUT
TNF-α responsive NF-κB/AP1-SEAP reporter assay
HEK-Blue™ TNF-α cells are designed to monitor human TNF-α-induced NF-κB/AP-1 stimulation or inhibition. This colorimetric bioassay can be used for screening activatory molecules, such as engineered cytokines, or inhibitory molecules, such as neutralizing antibodies.
HEK-Blue™ TNF-α cells respond specifically to recombinant human TNF-α . The reliable and consistent performance of HEK-Blue™ TNF-α cells makes them suitable for release assays of therapeutic molecules that inhibit TNF-α signaling, such as Adalimumab, a therapeutic monoclonal antibody targeting TNF-α (see figures).
Key features
- Readily assessable NF-κB/AP-1-SEAP reporter activity
- Convenient readout using QUANTI-Blue™ Solution
- High sensitivity to human (h) and mouse (m) TNF-α
- Stability guaranteed for 20 passages
Applications
- Therapeutic development
- Drug screening
- Release assay
Tumor necrosis factor alpha (TNF-α) is a multi-functional pro-inflammatory cytokine involved in regulating a wide spectrum of biological processes, such as cell proliferation, differentiation, and apoptosis.
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 TNF-α activity
hTNF-α EC50: 0.01 ng/ml (in medium) or 0.7 ng/ml (in water)
mTNF-α EC50: 0.1 ng/ml (in medium) or 3 ng/ml (in water)
Complete DMEM (see TDS)
Verified using Plasmotest™
Each lot is functionally tested and validated.
CONTENTS
Contents
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Product:HEK-Blue™ TNF-α Cells
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Cat code:hkb-tnfdmyd
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Quantity:3-7 x 10^6 cells
- 1 ml of Puromycin (10 mg/ml)
- 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)
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™ TNF-α cells were generated by stable transfection with the genes encoding for the human TNF-α receptor (TNFR1 and TNFR2 chains), as well as an NF-κB/AP-1-inducible SEAP secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of TNF-α to its receptor triggers a signaling cascade leading to NF-κB/AP1 activation and the subsequent production of SEAP. This can be readily assessed in the supernatant using QUANTI-Blue™ Solution, a SEAP detection reagent.
HEK-Blue™ TNF-α cells respond to recombinant human TNF-α, as well as recombinant mouse TNF-α (see figures). Of note, these cells are not responsive to human IL-1β. They can also be used for screening and release assay of molecules that inhibit TNF-α signaling, such as adalimumab, a monoclonal antibody targeting TNF-α.
TNF-α Background
Tumor necrosis factor-alpha (TNF-α) is a pleiotropic cytokine involved in necrotic and apoptotic cell death, cellular differentiation, inflammation, and regulation of immune cell activity [1]. Notably, deregulated TNF-α production has been implicated in a variety of conditions, including autoimmune and inflammatory diseases [1].
TNF-α is mainly produced by activated monocytes, macrophages, and T cells. It is first synthesized as a membrane-bound molecule that forms a compact homotrimer through non-covalent interactions. The trimeric membrane-bound form is cleaved by tumor necrosis factor-alpha converting enzyme (TACE) releasing the soluble trimer [2]. Both the membrane-bound and soluble TNF-α bind homotrimeric transmembrane receptors, TNFR1 or TNFR2, triggering signaling pathways that involve TRADD, TRAF2, and RIP, and leading to the activation of NF-κB and MAPK pathways.
1. Steeland S. et al., 2018. A new venue of TNF targeting. Int. J. Mol. Sci. 19:1442.
2. Brenner D. et al., 2015. Regulation of tumour necrosis factor signalling: live or let die. Nat Rev Immunol. 15(6):362-74.
DOCUMENTS
Documents
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
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