HEK-Blue™ IL-6 Cells

HEK-Blue™ IL-6 cells Unit size Cat. code Docs Qty Price
Bioactive human IL-6 sensor cells
3-7 x 10e6 cells

IL-6 Sensor Cells

HEK-Blue™ IL-6 cells allow the detection of bioactive human IL-6 by monitoring the activation of the STAT-3 pathway.  These cells were generated by stable transfection of HEK293 cells with the human IL-6R gene and a STAT3-inducible 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™. Stimulation of HEK-Blue™ IL-6 cells with recombinant human IL-6 can be blocked by anti-hIL-6-IgG.

APPLICATIONS: HEK-Blue™ IL-6 cells can be used to validate the functionality of recombinant native or engineered human IL-6. They can also be used to screen for molecules that inhibit IL-6 signaling such as antibodies targeting IL-6 or IL-6R and inhibitors of JAK1/JAK2.

Evaluation of IL-6 response in HEK-Blue hIL6 cells
Evaluation of IL-6 response in HEK-Blue hIL6 cells

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.

Evaluation of IL-6 inhibition in HEK-Blue hIL6 cells
Evaluation of IL-6 inhibition in HEK-Blue hIL6 cells

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.

Evaluation of specificity of HEK-Blue hIL6 cells
Evaluation of specificity of HEK-Blue hIL6 cells

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™.

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Antibiotic resistance: blasticidin, hygromycin, Zeocin™

Growth medium: DMEM, 4.5 g/l glucose, 2-4 mM L-glutamine, 10% (v/v) fetal bovine serum, 50 U/ml penicillin, 50 μg/ml streptomycin, 100 μg/ml Normocin™

Guaranteed mycoplasma-free

Shipped on dry ice

Specificity: human IL-6

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

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• 1 vial containing 3-7 x 106 cells

• 2 x 1 ml HEK-Blue™ Selection (250X concentrate); a solution containing several selection antibiotics.

• 1 ml Normocin™ (50 mg/ml)

• 1 pouch of QUANTI-Blue™ (SEAP detection medium)

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HEK-Blue IL6 pathway

The proinflammatory cytokine IL-6 is one of the most important mediators of fever and of the acute phase response. 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 the dimerization and translocation to the nucleus of STAT3 where it binds enhancer elements of IL-6-inducible genes.

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

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