Invitrogen™ CellSensor™ ISRE-bla Jurkat Cell Line

Description

The CellSensor™ ISRE-bla Jurkat cell line contains a beta-lactamase reporter gene under control of the interferon-stimulated response element (ISRE) stably integrated into Jurkat cells. To construct this cell line, the ISRE-bla construct was transduced into Jurkat cells by lentivirus. Flow cytometry was used to isolate cells responsive to IFNα. This cell line was validated for DMSO tolerance, cell number, stimulation time, substrate loading time, as well as Z' and EC50 concentrations of IFNα. The CellSensor™ ISRE-bla Jurkat cell line is responsive to Interferon alpha (IFN-alpha) and Interferon beta and can be used to probe the type I interferon-induced JAK-STAT signaling pathways. This cell line can be adapted for high-throughput screening of agonist or antagonist compound libraries. Candidate drugs can also be tested for dose response against this cell line. ISRE-bla Jurkat cells were treated with agonist IFN-alpha; over the indicated concentration range in a 384-well format. Cells were incubated for 5 hours with agonist and 0.5% DMSO and then combined with LiveBLAzer™-FRET B/G Substrate (CCF4-AM) for 2 hours. Fluorescence emission values at 460 nm and 530 nm were obtained using a standard fluorescence plate reader, and the 460/530 ratios were plotted against the concentration of the agonist.