Mammalian target of rapamycin (mTOR) is a serine/threonine protein kinase known to merge extracellular instructions with information about cellular metabolic resources and control the rate of anabolic and catabolic processes accordingly. In neurons, mTOR has been implicated in several aspects of development and physiology. The major problems with studying mTOR functions in neurons are caused by the restricted knowledge of downstream mTOR effectors and relatively poor tools for studying this particular kinase. In this chapter, we describe the materials and methods routinely used in our laboratory to study mTOR function and activity and the binding partners of mTOR in neurons.