Cytotoxic lymphocytes (CL) are highly motile cells that utilize granule exocytosis to kill virus-infected or transformed targets. Isolated CL and purified granule proteins have been used to investigate the molecular processes that CL use to kill their targets and to investigate the basis of human disease. We have set out various methods that are routinely used to isolate CL and characterize the cell death pathways they induce. As cell death mediated through TNF-superfamily members and their respective receptors is covered elsewhere, this manuscript will deal specifically with cytotoxic granule-mediated cell death.