Two-color coincidence single-molecule pulldown for the specific detection of disease-associated protein aggregates.

ABSTRACT: Protein misfolding and aggregation is a characteristic of many neurodegenerative disorders, including Alzheimer's and Parkinson's disease. The oligomers generated during aggregation are likely involved in disease pathogenesis and present promising biomarker candidates. However, owing to their small size and low concentration, specific tools to quantify and characterize aggregates in complex biological samples are still lacking. Here, we present single-molecule two-color aggregate pulldown (STAPull), which overcomes this challenge by probing immobilized proteins using orthogonally labeled detection antibodies. By analyzing colocalized signals, we can eliminate monomeric protein and specifically quantify aggregated proteins. Using the aggregation-prone alpha-synuclein protein as a model, we

SUBMITTER: Saleeb RS 

PROVIDER: S-EPMC10651132 | biostudies-literature | 2023 Nov

REPOSITORIES: biostudies-literature