HEK-Blue™ IL-6 Cells
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Human IL-6 Reporter Cells
3-7 x 10e6 cells
IL-6 Reporter 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 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™.
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.
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™.
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™
Specificity: human IL-6
Detection range: 0.03 - 10 ng/ml for human IL-6Back to the top
- 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)Back to the top
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.Back to the top