A significant proportion of genes in any prokaryotic or eukaryotic organism codes for proteins involved in transport processes. These carriers, receptors, and channels determine the flow of nutrients, ions, metabolites, and waste products across not just the plasma membrane but also, in eukaryotes, internal membranes—equilibrative transport when not energy-dependent and concentrative accumulation of substrate against a concentration gradient when assisted by energy expenditure of some kind. These gene products are not only essential for cellular functions and survival but can also serve as drug targets or determinants of drug uptake and extrusion. The protocol presented here can be utilized to study nutrient or drug uptake and/or efflux across the plasma membrane of kinetoplastid organisms.