IL-6 Reporter HEK 293 Cells

IL-6 Reporter Cells

HEK-Blue™ IL-6 Cells signaling pathway

HEK-Blue™ IL-6 cells allow the detection of bioactive human interleukin-6 (IL-6) by monitoring the activation of the STAT-3 pathway.  The proinflammatory cytokine IL-6 is one of the most important mediators of fever and of the acute phase response.

More details on proinflammatory cytokine IL-6

Cell line description:

HEK-Blue™ IL-6 cells were generated by stable transfection of human embryonic kidney  293 (HEK293)‑derived cells with the genes encoding human IL-6 receptor (IL-6R) and signal transducer and activator of transcription 3 (STAT3). In order to detect the activation of the IL-6 pathway, they were further transfected with a STAT3-inducible secreted embryonic alkaline phosphatase (SEAP) reporter gene.

Upon IL-6 stimulation, HEK-Blue™ IL-6 cells trigger the activation of STAT3 and the subsequent secretion of SEAP. Levels of STAT3-induced SEAP can be readily monitored using QUANTI-Blue™ Solution.

Stimulation of HEK-Blue™ IL-6 cells with recombinant human IL-6 can be blocked by anti-hIL-6-IgG.

Features of HEK-Blue™ IL-6 cells:

• Fully functional IL-6 signaling pathway
• Readily assessable SEAP reporter activity
• The stability for 20 passages has been validated
• Functionally tested and guaranteed mycoplasma-free

Applications of HEK-Blue™ IL-6 cells:

• Detection of human IL-6
• Screening of anti-IL-6 or anti-IL-6R antibodies
• Screening of JAK1/JAK2 inhibitors

Figures

Dose-response of HEK-Blue™ IL-6 cells to recombinant human IL‑6. Cells were stimulated with increasing concentration of recombinant human IL-6. After overnight incubation, the NF-κB response was determined using QUANTI‑Blue™, a SEAP detection reagent, and reading the optical density (OD)at 630 nm.

Dose-dependent inhibition of IL-6 response using Anti-hIL-6-IgG neutralizing antibody in HEK-Blue™ IL-6 cells. Anti-hIL-6-IgG was incubated with 1 ng/ml of hIL-6 for 30 minutes prior to the addition of HEK‑Blue™ IL-6 cells. After overnight incubation, SEAP activity was assessed using QUANTI‑Blue™. Data represent % inhibition of reporter activity.

Cytokine response profile of HEK-Blue™ IL-6 cells. Cells were stimulated with various human recombinant cytokines; 3 ng/ml IL-1β, IL-4, IL‑6, IL-13, IL-18, IFN-γ, TGF-β, TNF-α or 1x10⁴ IU/ml IFN-α and IFN-β. After overnight incubation, SEAP activity was assessed using QUANTI-Blue™.

Specifications

Antibiotic resistance: blasticidin, hygromycin, Zeocin®

Growth medium: DMEM, 4.5 g/l glucose, 2-4 mM L-glutamine, 10% (v/v) heat-inactivated fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 100 μg/ml Normocin™

Guaranteed mycoplasma-free

Specificity: human IL-6

Detection range: 0.03 - 10 ng/ml for human IL-6

This product is covered by a Limited Use License (See Terms and Conditions).

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 2 x 1 ml of HEK-Blue™ Selection (250x concentrate)
• 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)

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

Details

The proinflammatory cytokine IL-6 is one of the most important mediators of fever and of the acute phase response [1, 2].

IL-6 exerts its action by first binding to the IL-6R. The complex of IL-6 and IL-6R associates with the signal-transducing membrane protein gp130, thereby inducing its dimerization.

This leads to the activation by phosphorylation of the tyrosine kinases of the Janus family (JAK1, JAK2, and Tyk2).

Activated JAKs induce dimerization and translocation to the nucleus of STAT3 where it binds enhancer elements of IL-6-inducible genes [3].

In HEK-Blue™ IL-6 cells activation of the JAK/STAT3 pathway with IL-6 leads to the production of SEAP.

1. Kang S. et al., 2015. Therapeutic uses of anti-interleukin-6 receptor antibody. Int. Immunol. 27: 21-9.
2. Akira S. et al., 1990. Biology of multifunctional cytokines: IL 6 and related molecules (IL 1 and TNF). FASEB J. 4(11):2860-7.
3. Murray P., 2007. The JAK-STAT Signaling Pathway: Input and Output Integration. J. Immunol. 178:2623-9.

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