Control Reporter THP-1 Cells - THP1-Null Cells

THP1-Null cells are designed to study the signals involved in inflammasome activation. To become susceptible to inflammasome inducers, these cells must be induced by stimuli commonly used for induction in model systems, such as lipopolysaccharide (LPS) and phorbol 12-myristate acetate (PMA). Stimulation by LPS or differentiation with PMA induces the production of pro-IL-1β, the immature form of IL-1β. Subsequent stimulation with inflammasome inducers, such as ATP and alum, leads to caspase-1 activation and IL-1β maturation and secretion. Mature IL-1β can be detected by Western blot, ELISA, or a cell-based assay.

InvivoGen has developed a new method to detect bioactive IL-1β, based on HEK293 cells specifically engineered to selectively respond to IL-1β, named HEK-Blue™ IL-1β. These cells feature the SEAP (secreted embryonic alkaline phosphotase) reporter gene under the control of an NF-kB-inducible promoter. They naturally express the IL-1β receptor (IL-1R), and all the proteins involved in the MyD88-dependent IL-1R signaling pathway that leads to NF-kB activation. Thus upon IL-1β binding to IL-1R, a signaling cascade is initiated triggering NF-kB activation and the subsequent production of SEAP. Detection of SEAP in the supernatant of HEK-Blue™ IL-1β cells can be readily assessed using QUANTI-Blue™ Solution, a SEAP detection medium. QUANTI-Blue™ Solution turns blue in the presence of SEAP which can be easily quantified using a spectrophotometer.