Isabelle Vernos and a panel of hands-on experts present their most productive and reproducible techniques for the identification, purification, and characterization of the kinesin superfamily of microtubule-dependent motors. The methods range from the most basic to the most sophisticated and include step-by-step instructions and extensive cautionary notes to ensure experimental success. Among the approaches considered are methods to express and purify kinesins in different systems, to characterize microtubule-enhanced ATPase activity and motility properties, and to test microtubule destabilizing activity. Comprehensive and highly practical, Kinesin Protocols makes available all the key basic and cutting-edge methods needed to successfully study the multifaceted world of kinesin-like proteins and to explore their many functions.