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http://eprints.org/relation/isVersionOf https://researchdata.bbk.ac.uk/id/document/1031 http://eprints.org/relation/isVolatileVersionOf https://researchdata.bbk.ac.uk/id/document/1031 http://eprints.org/relation/issmallThumbnailVersionOf https://researchdata.bbk.ac.uk/id/document/1031 archive 126 disk0/00/00/01/90 2022-04-07 13:01:10 2026-04-14 11:12:21 2022-04-07 13:01:10 data_collection show MONISTROL Jim Department of Biological Sciences, Birkbeck College 0000-0002-4634-818X Saibil Helen Department of Biological Sciences, Birkbeck College 0000-0002-2266-8891 NS bcs public SDS PAGE, alpha-synuclein, HSP70, DNAJB1, Apg2 The dataset contains 1 file explaining the different conditions and 11 pictures of SDS PAGE showing the binding of various chaperones to alpha-synuclein fibres. 2022-04-04 Birkbeck College, University of London 10.18743/DATA.00190 yes ismb metadata_and_data_files yes Three independent repeats were performed for each condition. All chaperone aliquots (Hsc70, DNAJB1, Apg2) were centrifuged at 17,000 g for 30 minutes at 4°C. αSyn amyloid fibres were sonicated for 15 minutes at high frequency using a CPX 2800 Bransonic Ultrasonic bath (Branson). αSyn amyloid fibres (40 µM, monomer concentration) were incubated with Hsc70 (8 µM), DNAJB1 or ΔJ-DNAJB1 (4 µM), and Apg2 (different concentrations as indicated) in the disaggregation buffer for 1 hour at 30°C. The samples were then centrifuged for 45 minutes at 17,000 g to separate the pellet, containing the fibres and bound chaperones, from the supernatant with unbound chaperones. The supernatant was then collected and both fractions were incubated in 4X NuPAGE LDS Sample buffer (Thermo Fisher Scientific) for at least 30 minutes at 90°C. Samples were then loaded on BOLT 4-12% Bis-Tris gels (Thermo Fisher Scientific) and analysed by SDS-PAGE. Not applicable