Updated project metadata. We describe a single-step centrifugal elutriation method to produce synchronous G1-phase procyclic trypanosomes at a scale amenable for proteomic analysis of the cell cycle. Using ten-plex tandem mass tag technology, we quantified 5,325 proteins across the cell cycle in this parasite, providing a useful resource for the scientific community. Of these, 384 proteins were classified as cell cycle regulated and these were subdivided into nine distinct clusters of temporal regulation. A number of known cell cycle regulators in trypanosomes were detected in these groups, validating our approach, as well as forty novel and essential cell cycle regulated proteins that could be considered as future drug targets. Through cross-comparison to the TrypTag microscopy database, we were able to validate the cell cycle regulated patterns of expression for many of these proteins of unknown function. A convenient interface to access and interrogate these data is also presented.