HEK-Blue™ IFN-γ Cells

August 27^th 2019 - CONTENTS UPDATE NOTIFICATION
Please note that, for your convenience, these cells are now provided with QUANTI-Blue™ Solution.

Human Interferon-γ Reporter Cells

HEK-Blue™ IFN-γ cells allow the detection of bioactive human IFN-γ by monitoring the activation of the JAK/STAT-1 pathway.

HEK-Blue™ IFN-γ cells produce SEAP in response to IFN-γ stimulation only. They are unresponsive to type I IFNs.

Levels of SEAP in the supernatant can be easily determined with QUANTI-Blue™.

Figures

Cells were stimulated with increasing concentrations of hIFN-α (IU/ml), hIFN-β (IU/ml), or hIFN-γ (ng/ml). After a 24h incubation, the levels of IFN-induced SEAP were determined using QUANTI-Blue™, a SEAP detection reagent, and by reading the optical density (OD) at 655 nm.

Ligand	EC50	Response Ratio
Human IFN-γ	0.7 ng/ml	25

The response ratio was calculated by dividing the OD at 655 nm for the treated cells by the OD at 655 nm for the untreated cells.

Specifications

Antibiotic resistance: blasticidin, 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

Detection range for human IFN-γ: 5 - 100 IU/ml

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml of blasticidin (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)

Shipped on dry ice (Europe, USA & Canada)

Description

HEK-Blue™ 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-Blue™ IFN-γ cells produce SEAP in response to IFN-γ stimulation only. They are unresponsive to type I IFNs.

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.