Fig. 2: Verification of CSIC in plasma.
From: A novel approach to detecting plasma synuclein aggregates for Parkinson’s disease diagnosis
a Schematic representation of the experimental design. HC and PD plasma samples were subjected to αSyn depletion, followed by verification using b ELISA, c Western blotting, and d CSIC. b The αSyn-depleted supernatant was analyzed using sandwich ELISA to detect total αSyn. The red and blue bars represent PD and HC plasma, respectively. c αSyn was immunoprecipitated from HC and PD plasma samples using an anti-αSyn antibody (MJFR1). The presence of immunoprecipitated αSyn was verified by Western blotting using an anti-αSyn (211)-HRP antibody. The immunoprecipitated αSyn is indicated by an arrow. d CSIC was used to amplify αSyn aggregates in αSyn-depleted and untreated HC and PD plasma samples. The bar graph shows αSyn aggregates derived from CSIC. The red bar represents PD plasma, and the blue bar represents HC plasma. e CSIC was used to amplify αSyn aggregates in plasma samples from HCs (n = 3) and PD patients (n = 3). The red and blue lines represent amplified αSyn aggregates from PD and HC plasma, respectively. f Following CSIC, the final product was subjected to a sedimentation assay. The supernatant (sup) and pellet were separated, and equal volumes were analyzed using Western blotting. Immunoblotting was performed using an anti-αSyn (211)-HRP antibody. Arrows indicate the αSyn bands.
