TNF-α Dual Reporter HEK 293 Cells

NF-κB-SEAP & KI-[IL-8]-Lucia reporter cells 

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].

more More details

 

(*) 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

Target

TNF-α

Tested applications

Detection and quantification of human TNF-α activity

Cell type
Epithelial
Growth properties
Adherent
Tissue origin
Human embryonic kidney cells
Reporter gene
SEAP
Lucia®
Detection method
Colometric (SEAP), Bioluminescence (Lucia)
Detection range

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

Antibiotic resistance
Zeocin®
Growth medium

Complete DMEM (see TDS)

Mycoplasma-free

Verified using Plasmotest™

Quality control

Each lot is functionally tested and validated.

CONTENTS

Contents

  • Product: 
    HEK-Blue-Lucia™ TNF-α Cells
  • Cat code: 
    hkd-tnfa
  • Quantity: 
    3-7 x 10^6 cells
Includes:
  • 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
  • Storage:

    • Liquid nitrogen vapor
    Stability: 20 passages

    Caution:

    • Upon receipt, store immediately in liquid nitrogen vapor. Do not store cell vials at -80°C.

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

HEK-Blue-Lucia™ TNF-α Cells

Technical Data Sheet

Safety Data Sheet

Validation Data Sheet

Certificate of analysis

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