Optimized Measurements Of Transient Changes In Intracellular Calcium In Cells Following Agonist Activation with the CytoFLEX Analyzer

Generation of fluorescent antibody or genetic labels to identify hormone and neurotransmitter receptor activity can be difficult and time consuming. A useful alternative is recording physiological changes in response to agonist binding its cognate receptor, many of which are G-protein coupled. When an agonist binds a G-protein coupled receptor, it triggers a quick cascade of events that often results in a transient release of calcium from intracellular stores. This application note demonstrates an agonist-based calcium study using the peristaltic sample delivery of the new CytoFLEX analyzer. Using the ester based, green fluorescent calcium indicator, Fluo-4 AM, [Ca2+]i changes were measured in HEK-293 cells in response to ATP stimulation.