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.

Protocol

mRNA purification by translating ribosome affinity purification (TRAP) of DAT-expressing cells (dopaminergic neurons) in mouse ventral midbrain

This protocol describes the capture of eGFP-L10a-tagged ribosomes and mRNA from DAT-expressing cells (dopaminergic neurons) in mouse ventral midbrain.

Protocol

Differentiation of human medium spiny neurons (MSNs) from induced pluripotent stem cells (iPSCs)

This protocol generates human medium spiny neurons (MSNs) from induced human pluripotent stem cells (iPSCs).

Protocol

Differentiation of human cortical neurons (CNs) from induced pluripotent stem cells (iPSCs) and their coculture with rat astrocytes

This protocol described the production of human cortical neurons from induced pluripotent stem cells in cocultured with commercially available rat astrocytes.

Protocol

Quantification of extracellular 𝜶-synuclein assay

This protocol uses a classical ELISA experimental design with sensitive electrochemical detection to provide a measure of 𝜶-synuclein concentration in the conditioned media of cultured iPSC-derived human cells.

Protocol

Membrane and cytosol fractionation

This protocol describes membrane and cytosol fractionation of cells expressing different DNAJC5 isoforms.

Protocol

In vitro depalmitoylation assay

This protocol describes the in vitro depalmitoylation assay of DNAJC5.

Protocol

CRISPR/Cas9 genome editing

This protocol describes the generation of DNAJC5 KO using CRISPR/Cas9 edition.

Protocol

Medium fractionation and EV preparation

This protocol describes the characterization of the extracellular alpha-synuclein and DNAJC5.

Protocol

Co-immunoprecipitation

This protocol describes a common procedure to perform co-immunoprecipitation.

Protocol

Mitochondria purification

This protocol describes how to purify mitochondria from cell culture.

Protocol

Differentiation of SH-SY5Y cells

This protocol describes a standard procedure to differentiate SH-SY5Y cells into dopaminergic neurons using retinoic acid.

Protocol

shRNA knockdown

This protocol describes a standard procedure to generate a stable cell line for a DNAJC5 knockdown using shRNA.

Protocol

Immunoblots

The steps of this protocol are similar for different sources of protein such as cell lysate, cytosol, or membrane samples, and EV samples. However, some specific changes depend on the protein of interest, endogenous alpha-synuclein, or type of cells, hiPSC dopamine neurons.

Protocol

Fura-2 imaging of ionomycin response, with and without R568; a CaSR positive modulator

Fura-2 imaging protocol to accompany Kilfeather, Khoo et al., 2023: Single cell spatial transcriptomic and translatomic profiling of dopaminergic neurons in health, aging, and disease.