HEK-Blue™ IL-12 Cells

IL-12 Sensor Cells

HEK-Blue™ IL-12 cells are designed to detect bioactive human and mouse IL-12 by monitoring the activation of the STAT-4 pathway.  These cells were generated by stably introducing the human genes for the IL-12 receptor and the genes of the IL-12 signaling pathway into HEK293 cells. Furthermore, these cells express a STAT4-inducible SEAP reporter gene.

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 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™.

APPLICATIONS: HEK-Blue™ IL-12 cells can be used to validate the functionality of recombinant native or engineered human or mouse IL-12. They can also be used to screen for molecules that inhibit IL-12 signaling such as antibodies targeting IL-12 or IL-12R and inhibitors of TyK2/JAK2.

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™, 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
Recombinant hIL-13	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, 2-4 mM L-glutamine, 10% (v/v) fetal bovine serum, 50 U/ml penicillin, 50 μg/ml streptomycin, 100 μg/ml Normocin™

Guaranteed mycoplasma-free

Shipped on dry ice

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

• 100 μl Blasticidin (10 mg/ml)

• 100 μl Hygromycin B Gold (>98% pure hygromycin B; 100 mg/ml).

• 100 μl Zeocin™ (100 mg/ml)

• 1 ml Normocin™ (50 mg/ml)

• 1 pouch of QUANTI-Blue™ (SEAP detection medium)

Description

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). 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. 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.