Catalog
ASAP is committed to accelerating the pace of discovery and informing a path to a cure for Parkinson’s disease through collaboration, research-enabling resources, and data sharing. We’ve created this catalog to showcase the research outputs and tools developed by ASAP-funded programs.
Plasmid construction – A RAB7A Phosphoswitch coordinates Rubicon homology protein regulation of Parkin-dependent mitophagy
Plasmid construction via restriction enzyme digest and ligation.
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Mitophagy induction using Oligomycin/Antimycin A
Mitophagy induction in HeLa cells using Oligomycin/Antimycin A.
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Calculating mitochondrial protein solubility
Protocol for generating solubility calculations for mitochondrial proteins from starting ‘soluble’ and ‘insoluble’ fraction mass spectrometry data, using MaxQuant and Perseus software pipelines.
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Reverse-phase high pH fractionation (using Thermo Fisher, Cat# 84868)
This protocol uses the Pierce™ High pH Reversed-Phase Peptide Fractionation Kit (Thermo Fisher, Cat# 84868).
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Preparation of soluble and insoluble mitochondrial protein fractions for immunoblotting
Preparation of soluble and insoluble mitochondrial protein fractions from HeLa cells for immunoblotting.
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Mitochondrial isolation from HeLa cells
Mitochondrial isolation and quantification from HeLa cells.
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Western blotting using the BioRad Criterion Blotter system
Protocol for performing an SDS-PAGE Western blot analysis using the BioRad Criterion Blotter system.
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Preparing whole cell samples for immunoblot analysis
Protocol for preparation of HeLa cell lysates for immunoblot analysis.
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Mitophagy induction using Difereprone
Mitophagy induction in HeLa cells using Difereprone (DFP).
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