High quality screening by measurement of intracellular calcium levels.

摘要

A method for determining in vitro whether a candidate compound is an agonist or an inhibitor of a protein using cells that contain a sensitive messenger calcium comprising the steps of: (a) providing cells expressing the protein and contain a calcium sensitive messenger; (B) exposing the cells to a chelator of calcium under conditions in which the chelator to enter the cell and resides intracellularly; (C) exposing a first portion of the cells to a candidate compound and reserving a second portion of cells not exposed to the candidate compound; (D) measuring the signal generated in the first portion and the signal generated in the second portion of the cells; and (e) comparing the amount of signal generated in the first and second portions of the cells where an increase in the amount of signal in the presence of the candidate compound indicates that the candidate compound is an agonist of the protein. wherein the signal generated by the calcium sensitive messenger is retarded by the presence of calcium chelator within the cell and the protein is a G-protein coupled or an ion channel that is coupled to a calcium channel receptor.