IL-11 Reporter HEK 293 Cells

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Interleukin-11 Reporter Cells

Signaling pathway in HEK-Blue™ IL-11 cells

HEK-Blue™ IL-11 cells were engineered from the human embryonic kidney HEK 293 cell line to detect bioactive interleukin-11 (IL-11) by monitoring the activation of the JAK/STAT3 pathway. They can also be used for screening antibodies or small molecule inhibitors targeting the IL-11 pathway.

IL-11 has pleiotropic physiological functions, ranging from development and tissue homeostasis, to immune responses [1, 2].

 More details

Cell line description

HEK-Blue™ IL-11 cells were generated by stable transfection with the genes encoding the human IL-11 receptor (IL-11Rα and gp130 chains), human STAT3, and a STAT3-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of IL-11 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-11 cells detect human and murine IL-11 (see figures). Of note, these cells also respond to human IL-6 and IL-27 as they express the gp130 co-receptor shared by IL-11 and other cytokines of the IL-6 family (see figures).

Key Features

• Fully functional IL-11 signaling pathway
• Readily assessable STAT3-inducible SEAP reporter activity
• Strong response to human (h) and mouse (m) IL-11

Applications

• Detection and quantification of hIL-11 and mIL-11 activity 
• Screening of anti-IL-11 and anti-IL-11 receptor antibodies
• Screening of small molecule inhibitors of the IL-11 pathway

References:

1. Garbers C. & Scheller J., 2013. Interleukin-6 and interkeukin-11: Same same but different. Biol. Chem. 394:1145.
2. Kang S. et al., 2020. Historical overview of the interleukin-6 family cytokine. J. Exp. Med. 217: e20190347.

Figures

Human IL-11Rα mRNA expression in HEK-Blue™ IL-11 cells.
Total mRNA was extracted from ~1x10⁶ HEK-Blue™ parental cells (blue) and HEK-Blue™ IL-11 cells (red). Human IL-11Rα mRNA was amplified using quantitative RT-qPCR. Data are represented as the log2 fold change comparing hIL-11Rα relative expression between the two cell lines.

Dose-response of HEK-Blue™ IL-11 cells to recombinant IL-11.
Cells were stimulated with increasing concentrations of recombinant human IL-11 (hIL-11) and murine IL-11 (mIL-11). After overnight incubation, the STAT3 response was determined using QUANTI‑Blue™ Solution, a SEAP detection reagent. The optical density (OD) at 630 nm is shown as mean ± SEM.

Response of HEK-Blue™ IL-11 cells to a panel of cytokines.
Cells were stimulated with various human and murine recombinant cytokines: 1 ng/ml of hIL-11, mIL-11, or hIL-6, and 10 ng/ml hIL‑27 or TNF-α. After overnight incubation, SEAP activity was assessed using QUANTI‑Blue™ Solution. The optical density (OD) at 630 nm is shown as mean ± SEM.

Specifications

Antibiotic resistance: Blasticidin, Hygromycin B, and  Zeocin®

Growth medium: DMEM, 4.5 g/l glucose, 2 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-11 and mouse IL-11

Detection range:

• Detection range for human IL-11: 0.1 ng/ml - 100 ng/ml
• Detection range for murine IL-11: 0.1 ng/ml - 100 ng/ml

Quality Control:

• The expression of human IL-11Rα is confirmed by RT-qPCR.
• SEAP reporter activity in response to IL-11 is validated using functional assays.
• The stability for 20 passages following thawing is confirmed.
• These cells are tested for mycoplasma contamination. 

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml Normocin® (50 mg/ml)
• 2 x 1 ml of HEK-Blue Selection (250X)
• 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 cytokine interleukin 11 (IL-11) belongs to the IL-6 family,  which also includes IL-6, IL-27 p28/IL-30, IL-31, Leukemia inhibitory factor (LIF), Oncostatin M (OSM), Cardiotrophin-like cytokine (CLC), Ciliary neurotrophic factor (CNTF), Cardiotrophin-1 (CT-1), and Neuropoietin [1-3]. IL-11 has pleiotropic physiological functions, ranging from development and tissue homeostasis, to immune responses [1, 2]. IL-11 signalisation requires a multimeric structure comprising two specific α chains (IL-11Rα), two gp130 chains (common to the IL-6 signaling receptor), and 2 cytokines. This hexameric complex allows the juxtaposition of the intracellular Janus kinases (JAK) and activation of the signal transducer and activator of transcription 3 (STAT3) [1,3]. Other signaling pathways include the Ras-MAPK pathway, the PI3-K-Akt pathway, and the p38 and JNK MAPK pathways [2, 3].

While gp130 is ubiquitously expressed, IL-11Rα expression is cell-specific. Soluble IL-11Rα resulting from IL-11R proteolysis allows trans-signaling and widens the number of target cells [4]. Importantly, IL-11, like IL-6, has no affinity toward gp130 in the absence of its specific receptor α chain [1]. 

1. Garbers C. & Scheller J., 2013. Interleukin-6 and interkeukin-11: Same same but different. Biol. Chem. 394:1145.
2. Kang S. et al., 2020. Historical overview of the interleukin-6 family cytokine. J. Exp. Med. 217: e20190347.
3. Morris R. et al., 2018. The molecular details of cytokine signaling via the JAK/STAT pathway. Protein Science. 27:1984.
4. Lamertz L. et al., 2018. Soluble gp130 prevents interleukin-6 and interleukin-11 cluster signaling but not intracellular autocrine responses. Science Sign. 11(550):eaar7388).

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