Protocol Archive

This is a protocol for the preparation of Lentiviruses.

This is a protocol for human primary fibroblast culture.

This is a protocol for the direct conversion of human primary fibroblasts into neurons using a combination of transcription factor- and small molecule-based approach. The majority of converted neurons showing characteristics of cortical glutamatergic neurons.

This protocol is used to identify secreted inflammatory factors by cytokine profiling of the conditioned medium from human transdifferentiated neurons of healthy donors and AD patients.

This protocol is used for free-floating frozen (30-50 microns) and paraffin-embedded (10 microns) tissue sections.

This protocol describes the ex vivo fast-scan cyclic voltammetry (FSCV) technique for detecting dopamine in the dorsal striatum in mice.

These two protocols describe behavioral tests associated with Parkinson’s disease phenotypes in mice. The striatal motor learning protocol (rotarod test) is a two-phase paradigm assessing striatal motor learning under reversible dopamine antagonism.

Stereotaxic injection of viral vectors for gene knockdown experiments: The AAV vectors for knocking down the gene of interest were custom-made with Vector Biolabs using AAV-GFP-U6 vector for AAV1 packaging (Cat no, 7040, Vector Biolabs). They were then unilaterally injected into the striatum of mice as the protocol details.

Preparation of fresh paraformaldehyde for mouse perfusion and tissue fixation.

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.

The protocol described here is based upon previously established methods (Kinnebrew et al., 2019; Johnson and Radhakrishnan, 2021) and has also been used successfully to label cilia in RPE cells.

Preparation of soluble and insoluble mitochondrial protein fractions from HeLa cells for mass spectrometry analysis.

The authors investigated the colocalization of GFP-LRRK2 with a lysosomal localized TMEM192-3xHA.

This protocol details the generation of A549 PPM1H BromoTag CRISPR/CAS9 knock-in cell line.

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.

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

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

This protocol describes how to purify mitochondria from cell culture.

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

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