HEK-Dual™ IFN-γ Cells
|HEK-Dual™ IFN-γ cells||Unit size||Cat. code||Docs||Qty||Price|
Human IFN-γ SEAP/Luciferase Reporter Cells
3-7 x 10e6 cells
Human Interferon-γ Reporter Cells
HEK-Dual™ IFN-γ cells allow the detection of bioactive human IFN-γ by monitoring the activation of the JAK/STAT-1 pathway.
HEK-Dual™ IFN-γ cells were generated by stable transfection of HEK293 cells with the human STAT1 gene to obtain a fully active STAT1 pathway. The cells were further transfected with two reporter genes under the control of an IFN-γ inducible promoter.
As a result, HEK-Dual™ IFN-γ cells allow to study the activation of JAK/STAT-1 pathway, by monitoring the activity of two different reporters, SEAP or a secreted luciferase (Lucia).
HEK-Blue™ IFN-γ cells and HEK-Dual™ IFN-γ cells were incubated with increasing concentrations of recombinant human IFN-γ. After 24h incubation the levels of STAT1-induced SEAP and STAT1-induced Lucia luciferase were determined using QUANTI-Blue™ and QUANTI-Luc™, respectively. SEAP activity was assessed by measuring the OD at 655 while Lucia luciferase activity was determined by measuring Relative Light Units (RLU).
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™
- Detection range for human IFN-γ using QUANTI-Luc™: 1 - 10e3IU/ml
- Detection range for human IFN-γ using QUANTI-Blue™: 5 - 100IU/ml
These products are covered by a Limited Use License (See Terms and Conditions).Back to the top
- 1 vial of HEK-Dual™ IFN-γ Cells (3-7 x 10e6 cells)
- 1 ml of Blasticidin (10 mg/ml).
- 1 ml of Puromycin (10 mg/ml).
- 1 ml of Zeocin™ (100 mg/ml).
- 1 ml of 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)
- 1 pouch of QUANTI-Luc™ (Lucia luciferase detection medium)
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HEK-Dual™ IFN-γ cells were generated by stable transfection of HEK293 cells with the human STAT1 gene to obtain a fully active STAT1 pathway. The other genes of the pathway are naturally expressed in sufficient amounts.
The cells were further transfected with a SEAP reporter gene under the control of an ISG54 promoter fused to four interferon-gamma-activated sites (GAS).
HEK-Dual™ IFN-γ cells also feature the Lucia gene, a secreted luciferase reporter gene, under the control of the same inducible promoter that drives the SEAP reporter gene, that is an ISG54 promoter fused to four interferon-gamma-activated sites (GAS). As a result, HEK-Dual™ IFN-γ cells allow to study the activation of JAK/STAT-1 pathway, by monitoring the activity of two different reporters, SEAP or luciferase.
Simplified JAK/STAT signaling pathway induced by IFN-γ.
IFN-γ exerts its action by first binding to a heterodimeric receptor consisting of two chains, IFNGR1 and IFNGR2, causing its dimerization and the activation of specific Janus family kinases (JAK1 and JAK2). Two STAT1 molecules then associate with this ligand-activated receptor complex and are activated by phosphorylation. Activated STAT1 form homodimers and are translocated to the nucleus where they bind interferon-gamma-activated sites (GAS) in the promoter of IFN-γ inducible genes.Back to the top