IL-11 Reporter HEK 293 Cells

HEK-Blue™ IL-11 Cells signaling pathway

Interleukin-11 Reporter 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.  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 overexpression of the genes encoding the alpha chain of human IL-11 receptor (IL-11Rα), human STAT3, and a STAT3-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. STAT3-dependent SEAP activity is readily assessable in the supernatant using QUANTI-Blue™ Solution, a detection reagent. 
Of note, HEK-Blue™ IL-11 cells also respond to human IL-6 and IL-27 (see Figures) as they endogenously express the gp130 co-receptor shared by IL-11 and other cytokines of the IL-6 family.

Features of HEK-Blue™ IL-11 cells:

• Fully functional IL-11 signaling pathway
• Readily assessable STAT3-inducible SEAP reporter activity
• The stability for 20 passages has been verified
• Functionally tested and guaranteed mycoplasma-free

Applications of HEK-Blue™ IL-11 cells:

• Detection of human IL-11 (0.1 - 100 ng/ml) and murine IL-11 (0.1 -100 ng/ml)
• Screening of anti-IL-11 and anti-IL-11Rα antibodies

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, 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: human & murine IL-11

Detection range: 

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

Quality Control:

• The expression of human IL-11Rα has been confirmed by RT-qPCR.
• STAT3-dependent SEAP reporter activity in response to human and murine IL-11 has been validated.
• The stability for 20 passages, following thawing, has been verified. 
• These cells are guaranteed mycoplasma-free. 

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

Contents

• 3-7 x 10⁶ HEK-Blue™ IL-11 cells in a cryovial or shipping flask
• 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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