Jul 28, 2025
Version 2
Preparation of α-synuclein pre-formed fibrils (PFF) V.2
- Zoe Manglano-Artuñedo1,
- Jara Montero-Muñoz2,3,
- Antonella Consiglio2,3,
- Salvador Ventura1,
- Miquel Vila4
- 1Institut de Biotecnologia i de Biomedicina and Departament de Bioquímica i de Biologia Molecular, Universitat Autònoma de Barcelona, 08193 Bellaterra (Barcelona), Spain.;
- 2Department of Pathology and Experimental Therapeutics, Bellvitge University Hospital- IDIBELL, 08908 Hospitalet de Llobregat, Spain;
- 3Institute of Biomedicine of the University of Barcelona (IBUB), Carrer Baldiri Reixac 15-21, Barcelona 08028, Spain.;
- 4VHIR-CIBERNED-ASAP
- Vilalab Public
Protocol Citation: Zoe Manglano-Artuñedo, Jara Montero-Muñoz, Antonella Consiglio, Salvador Ventura, Miquel Vila 2025. Preparation of α-synuclein pre-formed fibrils (PFF). protocols.io https://dx.doi.org/10.17504/protocols.io.14egnrzk6l5d/v2Version created by Miquel Vila
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: July 28, 2025
Last Modified: July 28, 2025
Protocol Integer ID: 223412
Keywords: synuclein
Abstract
Protocol for preparation of α-synuclein pre-formed fibrils (PFF) obtained from expression and purification of Human α-synuclein
Troubleshooting
α- synuclein was dissolved in PBS buffer and placed in 96 wells plates containing teflon
polyball (1/8′′ diameter)
Plates were fixed into an orbital culture shaker Max-Q 4000 Thermo Scientific (Waltham, MA, USA) to keep the incubation at 37 °C, 100 rpm for 24 hours.
After production, PFF were briefly sonicated and added to the co-cultures for 8 hours at a concentration of 10ug/ml