IL-12 Reporter HEK 293 Cells

IL-12 Reporter Cells

HEK-Blue™ IL-12 Cells signaling pathway

HEK-Blue™ IL-12 cells are designed to detect bioactive human and murine interleukin-12 (IL-12) by monitoring the activation of the STAT-4 pathway.

IL-12 is a pro-inflammatory cytokine that regulates T-cell and natural killer-cell responses, as well as inducing the production of interferon-γ (IFN-γ) [1-3].

 More details

Cell line description:

HEK-Blue™ IL-12 cells were generated by stably introducing the human genes for the IL-12 receptor and the genes of the IL-12 signaling pathway into the human embryonic kidney HEK 293 cell line. Furthermore, these cells express a STAT4-inducible SEAP reporter gene.

The binding of IL-12 to the IL-12R on the surface of HEK-Blue™ IL-12 cells triggers a signaling cascade leading to the activation of STAT-4 with the subsequent production of SEAP.

Detection of SEAP in the supernatant of HEK-Blue™ IL-12 cells can be readily assessed using QUANTI-Blue™ Solution.

Features of HEK-Blue™ IL-12 cells:

• Fully functional IL-12 signaling pathway
• Readily assessable SEAP reporter activity
• Functionally tested and guaranteed mycoplasma-free

Applications of HEK-Blue™ IL-12 cells:

• Detection of human and murine IL-12
• Screening of anti-IL-12 and anti-IL-12R antibodies
• Screening of TyK2/JAK2 inhibitors

Figures

Response of HEK-Blue™ IL-12 cells to increasing concentrations of human and murine recombinant IL-12.

After a 24h incubation, the levels of SEAP were determined using QUANTI-Blue™ Solution,  a SEAP detection reagent, and by reading the optical density (OD) at 630 nm.
 

Ligand	EC50	Response Ratio
Human IL-12	4.2 ng/ml	13
Murine IL-12	4.1 ng/ml	11

 

The response ratio was calculated by dividing the OD at 630 nm for the treated cells by the OD at 630 nm for the untreated cells.

 

Specifications

Antibiotic resistance: Blasticidin, Hygromycin, 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™

Guaranteed mycoplasma-free

Specificity: human and mouse IL-12

Detection range: 1 - 100 ng/ml for human and mouse IL-12

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 2 x 1 ml HEK-Blue™ Selection (250x concentrate)
• 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 12 (IL-12), formerly known as cytotoxic lymphocyte maturation factor (CLMF) and natural killer cell stimulatory factor (NKSF), is primarily produced by dendritic cells and macrophages.

It promotes the development of Th1 responses and is a powerful inducer of interferon-gamma (IFN-γ) [1-3].

IL-12 exerts its biological effect following binding to the IL-12 receptor (IL-12R), which is a heterodimeric receptor composed of two subunits, IL-12R-β1 and IL-12R-β2.

The binding of IL-12 to its receptor triggers a signaling pathway involving TyK2 (tyrosine kinase 2), JAK2 (Janus kinase 2), and STAT4 (signal transducer and activator of transcription 4) which results in the production of IFN-γ.

In HEK-Blue™ IL-12 cells, activation of the STAT4 pathway with IL-12 leads to the production of SEAP.

1. Teng M.W. et al., 2015. IL-12 and IL-23 cytokines: from discovery to targeted therapies for immune-mediated inflammatory diseases. Nat Med. 21(7):719-29.
2. Vignali DA. & Kuchroo VK., 2012. IL-12 family cytokines: immunological playmakers. Nat Immunol. 13(8):722-8.
3. Trinchieri G., 2003. Interleukin-12 and the regulation of innate resistance and adaptive immunity. Nat Rev Immunol. 3(2):133-46.

FAQ

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