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Untargeted lipidomics of Tagless Lyso-IP
onLysosomal biology is implicated in neurodegenerative diseases and health. It has traditionally been difficult to profile the lipidolomic homeostasis of the lysosome in disease states. To overcome this challenge, the authors have developed the Tagless Lyso-IP method to rapidly prepare lysosome-enriched samples from human peripheral blood.
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Cathepsin D assay to verify the retention of lysosomal content
onCathepsin D assay is a fluorescence-based assay that leverages the activity of cathepsin D to monitor the intactness of lysosomes in the cell. Here, the authors describe a method where the measurement of cathepsin D activity is used to verify the intactness of lysosomes.
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Sample preparation protocol for proteomic analysis of isolated lysosomes and whole cell extracts
onHere, the authors describe a step-wise protocol for proteomic sample preparation derived from cell line models or isolated human cells. The protocol has been optimized for organelle pulldown preps.
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Fixation and immunostaining protocol
onFixation and immunostaining protocol for tissue and cell culture.
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Tagless LysoIP method for molecular profiling of lysosomal content in clinical samples
onPreprint: Profiling lysosomal content using tag-based lysosomal immunoprecipitation (LysoTagIP) in cell and animal models allowed major discoveries, however studying lysosomal dysfunction in human patients remains challenging. Here, the authors report the development of the tagless LysoIP method to enable rapid enrichment of lysosomes from clinical samples and human cell lines.
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MS proteomics data related to “Disruption of lysosomal proteolysis in astrocytes facilitates midbrain organoid proteostasis failure in an early-onset Parkinson’s disease”
onMS proteomics data related to "Disruption of lysosomal proteolysis in astrocytes facilitates midbrain organoid proteostasis failure in an early-onset Parkinson’s disease" deposited to the MassIVE repository with the dataset identifier MSV000090202.
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Metabolomics data related to “Disruption of lysosomal proteolysis in astrocytes facilitates midbrain organoid proteostasis failure in an early-onset Parkinson’s disease”
onThe metabolomics is submitted at the NIH Common Fund’s National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org where it has been assigned Project ID (PR001491). The data can be accessed directly via its Project DOI: (https://doi.org/10.21228/M80M7R).
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Analysis codes related to “Disruption of lysosomal proteolysis in astrocytes facilitates midbrain organoid proteostasis failure in an early-onset Parkinson’s disease model”
onAnalysis codes related to "Disruption of lysosomal proteolysis in astrocytes facilitates midbrain organoid proteostasis failure in an early-onset Parkinson’s disease model."