IL-6 Reporter HEK 293 Cells
Product | Unit size | Cat. code | Docs. | Qty. | Price | |
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HEK-Blue™ IL-6 cells Human IL-6 Reporter Cells |
Show product |
3-7 x 10e6 cells |
hkb-hil6
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HEK-Blue™ IL-6 vial Additional cell vial |
Show product |
3-7 x 10e6 cells |
hkb-hil6-av
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Notification: Reference #hkb-hil6-av can only be ordered together with reference #hkb-hil6.
Interleukin-6 Reporter Cells
Signaling pathway in HEK-Blue™ IL-6 cells
HEK-Blue™ IL-6 cells were engineered from the human embryonic kidney HEK 293 cell line to detect bioactive human interleukin-6 (IL-6) by monitoring the activation of the JAK/STAT3 pathway. They can also be used for screening antibodies or small molecule inhibitors targeting the IL-6 pathway.
IL-6 is a multifunctional cytokine that plays a key role in the regulation of the immune response, hematopoiesis, and inflammation. It is one of the most important mediators of fever and of the acute phase response [1, 2].
Cell line description
HEK-Blue™ IL-6 cells were generated by stable transfection with the genes encoding the IL-6Rα chain of the human IL-6 receptor, as well as human STAT3 to obtain a fully active IL-6 signaling pathway. The gp130 chain of the IL-6 receptor is endogenously expressed. They were further transfected with a STAT3-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-6 to its receptor triggers a signaling cascade leading to the activation of STAT3 and the subsequent production of SEAP. This can be readily assessed in the supernatant using QUANTI-Blue™ Solution, a SEAP detection reagent.
HEK-Blue™ IL-6 cells detect human IL-6 (see figures). Of note, these cells also exhibit a moderate response to human IFN-γ (see figures). Stimulation of HEK-Blue™ IL-6 cells with recombinant human IL-6 can be blocked by anti-hIL-6-IgG (see figures).
Key features
- Fully functional IL-6 signaling pathway
- Readily assessable STAT3-inducible SEAP reporter activity
- Strong response to human IL-6
Applications
- Detection and quantification of hIL-6 and mIL-6 activity
- Screening of anti-IL-6 and anti-IL-6 receptor antibodies
- Screening of small molecule inhibitors of the IL-6 pathway
References:
1. Johnson DE. et al., 2018. Targeting the IL-6/JAK/STAT3 signalling axis in cancer. Nat Rev Clin Oncol. 15(4):234.
2. Hunter CA. & Jones SA., 2015. IL-6 as a keystone cytokine in health and disease. Nat Immunol. 16(5):448.
Specifications
Antibiotic resistance: Blasticidin, Hygromycin B, 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®
Specificity: Detects human IL-6
Detection range:
- Detection range for human IL-6: 0.03 ng/ml - 10 ng/ml
Quality Control:
- SEAP reporter activity in response to IL-6 is validated using functional assays.
- The stability for 20 passages following thawing is confirmed.
- These cells are tested for mycoplasma contamination.
This product is covered by a Limited Use License (See Terms and Conditions).
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- 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.