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Identification of compounds that affect cell movement is crucial to drug discovery. Drug components may have unexpected consequences with respect to cell motility, which would exclude these compounds in drug development. Finally, the development of drugs that target chemotactic pathways may be useful in the treatment of tumors, which often reprogram chemotactic pathways to become metastatic. The effects of these agents on cell movement can be measured using several different in vitro chemotactic assays. This review details the procedures of three in vitro measurements of chemotaxis: the Boyden chamber, the under\u2010agarose assay, and the automated, real\u2010time, ECIS\/Taxis assay, and discusses the inferences that can be drawn from the results of such studies. <jats:italic>Curr. Protoc. 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