Objective

Gq protein-coupled receptor activation is commonly monitored in live cells in real time using calcium-sensitive dyes and a fluorescence plate reader. Automated liquid handling within the plate reader is generally preferred to deliver agonists to the cells in the microplate while the detection system takes real-time readings of compound-induced changes in fluorescence intensity values. Analysis of the resulting kinetic readings yields information about the compound response profiles, including EC50 and IC50 values for agonists and antagonists.

In this poster we describe how the SpectraMax iDx Multi-Mode Microplate Readers with the SpectraMax Injector System featuring SmartInject™ Technology can be used to perform calcium assays on two different cell lines. 1321N1 (ECACC 86030402) is an adherent human astrocytoma cell line, expressing endogenous muscarinic cholinergic receptors.  THP 1 (ECACC 88081201) is a non-adherent human monocytic leukaemia cell line endogenously expressing purinergic P2Y2 receptors

Data analysis performed using SoftMax Pro Software yielded consistent EC50 and IC50 values for both cell lines, results were comparable with data generated on the FlexStation 3 Multi-Mode Microplate Reader.