IL-2 Reporter HEK 293 Cells

Human & Murine IL-2 Reporter Cells

HEK-Blue™ IL-2 Cells signaling pathway

HEK-Blue™ IL-2 reporter cells have been specifically designed to monitor the activation of the JAK-STAT pathway by both human and murine IL-2.

These cells were generated by stable transfection of the human embryonic kidney HEK 293 cell line with the human IL-2Rα, IL-2Rβ, and IL-2Rγ genes, along with the human JAK3 and STAT5 genes to obtain a fully functional IL-2 signaling pathway. In addition, a STAT5-inducible SEAP reporter gene was also introduced.

Upon IL-2 stimulation, HEK-Blue™ IL-2 cells trigger the activation of STAT5 and the subsequent secretion of SEAP. The levels of STAT5-induced SEAP can be readily monitored using QUANTI-Blue™ Solution.

HEK-Blue™ IL-2 cells can be used to validate the functionality, toxicity, and variable dosage effects of human or murine IL-2.

FEATURES OF HEK-BLUE™ IL-2 CELLS:

• Fully functional IL-2 signaling pathway
• Readily assessable SEAP reporter activity
• Functionally tested and guaranteed mycoplasma-free

APPLICATIONS OF HEK-BLUE™ IL-2 CELLS:

• Detection of human and murine IL-2
• Screening of anti-IL-2 antibodies

Read our review on IL-2: The Activator and Controller

Figures

Cells were stimulated with increasing concentration of recombinant human and murine IL-2. After overnight incubation,  the  STAT5  response was determined using QUANTI-Blue™ Solution, a SEAP detection reagent, and reading the optical density (OD) at 630 nm. EC₅₀ values are shown in the table above as mean ± SD.

Cells were stimulated with various human recombinant cytokines; 1 ng IL-2 and 10 ng/ml IL-1β, IL-4, IL-6, IL-18, TGF-β, TNF-α, IFN-γ, and 1x10⁴ IU/ml IFN-α, IFN-β. After overnight incubation, SEAP activity was assessed using QUANTI-Blue™ Solution.

Specifications

Antibiotic resistance: blasticidin, hygromycin, puromycin, Zeocin™

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

Guaranteed mycoplasma-free

Specificity: human & murine IL-2

Detection range: 0.03 - 10 ng/ml for human & murine IL-2

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml puromycin (10 mg/ml)
• 2 x 1 ml HEK-Blue™ CLR Selection (250X concentrate); a solution containing several selection antibiotics.
• 1 ml 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)

Details

Interleukin-2 (IL-2), also known as T cell growth factor, is mainly produced by activated T lymphocytes following antigen stimulation. This secreted cytokine plays an important role in immune activation and homeostatis [1,2]. Specifically, IL-2 plays dual roles by inducing natural killer cells as well as promoting the differentiation of naïve CD4+ T cells into regulatory T cells (Treg) cells. Interestingly, IL-2 has been approved for use as a high‑dose immunotherapeutic agent against cancer [1]. Additionally, research indicates that IL-2 has potential as a low-dose therapy for autoimmune diseases.
IL-2 exerts its action by binding to the heterotrimeric IL-2 receptor (IL‑2R), consisting of IL-2Rα (CD25), IL-2Rβ (CD122), and the common interleukin chain, IL-2Rγ (CD132). This binding leads to the activation of a signaling cascade through tyrosine kinases of the Janus family (JAK1 and JAK3) and signal transducer and transcription activator 5 (STAT5), which initiates transcriptional elements and regulates the expression of IL-2-inducible genes.

1. Choudhry H. et al., 2018. Prospects of IL-2 in Cancer Immunotherapy. Biomed Res Int. 2018:9056173.
2. Boyman O. & Sprent J., 2012. The role of interleukin-2 during homeostasis and activation of the immune system. Nat Rev Immunol. 12(3):180-90.

FAQ

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