Parkinson's disease (PD) is multifactorial, likely resulting from an intricate relationship of genetic and environmental factors affecting fundamental cellular processes. Histopathological hallmarks of PD include the development of granular inclusions known as Lewy bodies that are enriched with aggregates of the protein alpha-synuclein (alpha S). Historically, alpha S has been considered a natively unfolded protein prone to amyloidogenic behavior. However, recent studies have revealed a physiologically relevant folded alpha S tetramer that is both alpha-helical and aggregation-resistant. The two forms are thought to reside in a dynamic coexistence within cells, and it has been suggested that a shift from metastable tetramers to the monomeric form could serve as a mechanism for disease initiation. The underlying pathology causing this type of shift remains unknown, but the importance of understanding tetramer stability and disassembly has therapeutic potential that cannot be overemphasized. Isolation of tetrameric alpha S is complicated by its dynamic nature, so thorough and detailed biochemical and biophysical studies on this alpha S conformer have been hampered by accessibility issues. We now report a robust and reliable recombinant expression platform that enables purification of native tetrameric alpha S without any detergents or other structure-modifying additives.