Output 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.
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Output Type
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all protocols related to “Spermidine suppresses glial inflammation and parkinsonian abnormalities in ATP13A2 deficiency”
All protocols related to "Spermidine suppresses glial inflammation and parkinsonian abnormalities in ATP13A2 deficiency"
Generation of stable cell lines (Lentiviral vectors)
Generation of stable cell lines.
in vivo TurboID
Protocol uses iBioID with Lamp1-TurboID in astrocytes to study endo-/lysosomal protein interactions in mice. AAV delivery allows selective biotinylation for proteome mapping, offering insights into astrocyte signaling in vivo.
Postnatal astrocyte labeling by electroporation
Here, we provide a step-by-step guide to perform Postnatal Astrocyte Labeling by Electroporation (PALE), detailing the preparation, electroporation procedure, and post-electroporation analysis.
IHC staining (DAB) of Iba1 and Gfap
This protocol details the IHC staining (DAB) of Iba1 and Gfap.
Immunocytochemistry – subcellular localization of ATP13A4
This protocol describes an optimized method for visualizing the subcellular localization of over-expressed ATP13A4 in HeLa cells using immunocytochemistry.
Protein quantification by SDS-PAGE and Coomassie staining
This protocol provides a step-by-step guide for quantifying protein concentration by SDS-PAGE and Coomassie staining.
Expression and purification of ATP13A4 from Saccharomyces cerevisiae
This protocol describes the expression and purification of human ATP13A4 with a C-terminal Twin-Strep tag from Saccharomyces cerevisiae using a CuSO₄-inducible system.
Immunoblotting protocol
This protocol outlines a step-by-step method for performing western blot analysis, including protein separation by gel electrophoresis, transfer to membranes, antibody incubation, and detection.
Measurement of ATP13A4 ATPase activity using ADP-Glo Max Assay
This protocol describes a luminescence-based ADP-Glo Max assay (Promega) to measure the ATPase activity of ATP13A4 by detecting the rate of ATP hydrolysis.
Purification of ATP13A4 from HEK293T cells
This protocol describes the purification of ATP13A4 from HEK293T cells stably overexpressing ATP13A4. The procedure includes cell lysis, membrane solubilization, and affinity purification using Strep-TactinXT resin.
Auto-(de)phosphorylation assay
Protocol measures ATP13A4 phospho-enzyme levels by detecting its phosphorylated intermediate on an aspartate residue. Utilizes SDS-PAGE under acidic conditions and autoradiography to detect [32P]-labeled intermediates.
Fluorescently labeled polyamine uptake (flow cytometry)
Assess polyamine uptake capacities of a specific cell line after incubation with fluorescently labeled polyamines and mean fluorescence intensitiy acquisition via flow cytometry.
Fluorescently labeled polyamine uptake (confocal microscopy)
Method visualizes polyamine uptake in cells using fluorescently labeled polyamines and confocal microscopy.
RT-qPCR (cell lines)
This protocol describes the use of RT-qPCR to assess msAtp13a4 mRNA expression following lentiviral knockdown in C8-D1A cells.
