Sep 29, 2025

Public workspaceAutophagic flux analysis

DOI
https://dx.doi.org/10.17504/protocols.io.eq2ly49pplx9/v1
  • Maria Jose Perez J.1
  • 1Institut Imagine
  • Team Deleidi
Federico Bertoli
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DOI: https://dx.doi.org/10.17504/protocols.io.eq2ly49pplx9/v1
Protocol Citation: Maria Jose Perez J. 2025. Autophagic flux analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.eq2ly49pplx9/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: September 25, 2025
Last Modified: September 29, 2025
Protocol Integer ID: 228171
Keywords: autophagic flux analysis autophagic flux analysis
Funders Acknowledgements:
ASAP
Abstract
Autophagic flux analysis
Troubleshooting
Treat iPSC-derived microglia with 100 nM bafilomycin A1 (Sigma-Aldrich) for 4 hours prior to protein extraction.
Detect LC3B levels by Western blot and quantify the intensity of the LC3-II band using ImageJ.
Calculate autophagic flux as the ratio of LC3-II in bafilomycin-treated cells versus untreated cells.