Jun 04, 2025

Synaptosome extraction

DOI
https://dx.doi.org/10.17504/protocols.io.5qpvowmb7l4o/v1
  • Lucas Blasco-Agell1,2,
  • Meritxell Pons-Espinal1,2,
  • Antonella Consiglio1,2,
  • Miquel Vila3
  • 1Department of Pathology and Experimental Therapeutics, Bellvitge University Hospital- IDIBELL, 08908 Hospitalet de Llobregat, Spain;
  • 2Institute of Biomedicine of the University of Barcelona (IBUB), Carrer Baldiri Reixac 15-21, Barcelona 08028, Spain;
  • 3VHIR-CIBERNED-ASAP
  • Vilalab Public
Miquel Vila
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DOI: https://dx.doi.org/10.17504/protocols.io.5qpvowmb7l4o/v1
Protocol Citation: Lucas Blasco-Agell, Meritxell Pons-Espinal, Antonella Consiglio, Miquel Vila 2025. Synaptosome extraction. protocols.io https://dx.doi.org/10.17504/protocols.io.5qpvowmb7l4o/v1
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: June 04, 2025
Last Modified: June 04, 2025
Protocol  Integer ID: 219564
Keywords: synaptosome extraction
Funders Acknowledgements:
ASAP
Grant ID: ASAP-020505
Abstract
Synaptosome extraction
Synaptosomes (SYNs) are then extracted from mature iPSC-derived cortical neurons using Syn-PER‱ Synaptic Protein Extraction Reagent (Thermo Fisher Scientific).
1x of protease inhibitors (Roche) are added to the Syn-PER‱ Reagent immediately before use. 200 μL of Syn-PER‱ Reagent was added to each 6-wp well and cells were lifted from the surface using a cell scraper (Biologix).
Samples are centrifuged at 1200xg for 10 minutes at 4ºC.
Supernatant is transferred to a new tube and centrifuged at 15000xg for 20 minutes at 4ºC. Pellet containing SYNs was resuspended in Syn-PER‱ Reagent.
SYNs concentration is measured using the Bradford assay solution (Bio-Rad Laboratories)
Protein concentration is quantified by optical density in a PowerWave‱ XS multiplate reader (Biotek‱).
Resulting SYNs are frozen in 5% (v/v) Dimethyl sulfoxide (DMSO) at -80 ºC for extended periods, until pHRodoTM Red (ThermoFisher Scientific) conjugation.