An In Vitro Microplate Assay That Can Monitor IL-6 Secretions From SKOV-3 Ovarian Carcinoma Cells

By Brad Larson and Peter Banks, BioTek Instruments, Inc., Nicolas Pierre, CisBio US Inc., and Stephanie Georgiou, Enzo Life Sciences

Using the Cytation™3 Cell Imaging Multi-Mode Reader to Monitor IL-6 Secretion Using HTRF Detection and Cell Viability Using Digital Widefield Fluorescence Microscopy

Epidermal growth factor receptor (EGFR) is expressed in up to 70% of epithelial ovarian cancers (EOCs). High levels of IL-6 have also been found in the ascites of EOC patients. It has been shown that ligand stimulated EGFR activated NFkB dependent transcription and induced secretion of the pro-inflammatory IL-6 cytokine. In this work we will demonstrate a multiplexed assay for inhibitors at the level of EGFR and NFkB to monitor IL-6 secretion and cell viability. The work flow of the multiplexed assay uses an assay plate and HTRF detection plate. Following treatment of the cells, supernatant is transferred to the HTRF detection plate where IL-6 concentrations are determined; then cell viability is assessed in the assay plate using fluorescent probes and imaging. All assays were conducted on the Cytation™3 Cell Imaging Multi-Mode Reader.