Here, we describe a detailed protocol to set up the αS-PMCA assay using αS synthetic aggregates in buffer and to accurately detect endogenous αS aggregates from human CSF samples. Given the amplificative nature of the technique, minute amounts of misfolded protein aggregates circulating in human bodily fluids can be multiplied and thereafter detected by more conventional methods, such as immune assays or fluorescence. Following these principles, αS-PMCA was standardized for the highly sensitive and specific detection of αS misfolded aggregates in cerebrospinal fluid (CSF) of patients with synucleinopathies.
Keywords: PMCA; Parkinson’s; Synucleinopathies; α-Synuclein.