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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Program
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CRN Team Name
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Theme
Protocols Suite — SH-SY5Y / iCell® Neurons / High-Content Imaging
Maintenance of SH-SY5Y cells with SNCA overexpression in DMEM/F-12, differentiation with RA/BDNF, and culture of iCell DopaNeurons with SNCA mutation. Treatment and immunostaining for downstream assays using specific protocols and analysis tools.
Immunofluorescencent Staining of A-synuclein, TFEB, GCase and LAMP1 in treated cultured cells
Protocol used to detect a-synuclein, TFEB, GCase, and LAMP1 via immunofluorescence in SH-SY5Y cells with mutant SNCA gene.
GCase activity assay
Protocol measures GCase activity using 4-MUG substrate. GCase protein from a cellular model is diluted in acidic buffer to mimic lysosome pH. Adding lipid/detergent maintains enzyme activity. Taurocholate bile salt is included in the protocol.
Protocol — DQ™ Red BSA Lysosomal Activity Assay HCS Analysis
Lysosomal proteolysis can be measured using self-quenched DQ™ Red BSA, followed by confocal imaging for spot-based quantification.
Protocol — LysoTracker® Red DND-99 Live-Cell Staining 26 HCS Analysis
Text summary: Live-cell lysosomal staining done with LysoTracker Red DND-99, followed by high-content confocal imaging using Opera Phenix Plus, and automated quantification in Harmony/Columbus.
Protocol — Quantitative Real-Time PCR (qRT-PCR): 3K-SNCA cells
Gene expression in 3K-SNCA SH-SY5Y cells was analyzed using Qiagen RNA extraction and TaqMan qRT-PCR on QuantStudio 1 platform for differentiated and undifferentiated cells.
Processing Serum, Plasma, Plasma EDTA from Whole Blood
Protocols cover serum, plasma, and plasma EDTA processing from whole blood samples.
Stool Sample Collection Protocol
The protocol outlines procedures for stool sample collection.
Isolation of Classical Monocytes from PBMCs (MB Kit No. 130-117-337)
Protocol isolates classical monocytes from peripheral blood mononuclear cells.
PBMCs isolation with CT Tubes (8mL Whole blood)
Protocol isolates peripheral blood mononuclear cells using CPT tubes.
Isolation of Platelets Protocol
The text describes a platelet isolation protocol.
Indirect Proximity Ligation Assay (PLA) – Brightfield
The authors describe the PLA protocol that is routinely used in the laboratory to detect nitrated alpha-synuclein and nitration of mitochondrial enzymes such as SOD2 and the mitochondrial complex 1 subunit NDUFB8.
qPCR of α-synuclein, TNF and NF-κβ
A procedure for quantitative real time reverse transcription PCR of α-synuclein, TNF, and NF-κβ.
STC-1 cell culture
Cell culture of STC-1 mouse enteroendocrine cells.
Immunohistochemistry using paraffin embedded tissue
Immunohistochemistry using paraffin embedded tissue
