Protocol Archive

Purification and Crystallization of ATG9 HDIR (828-839) fused ATG101 (1-198):ATG13 (1-197).

This protocol describes the process of expressing and purifying the nicking Cas9-MMLV RT fusion protein for prime editing.

In this protocol, we have listed the steps to purify the Vps13 adaptor binding domain (or VAB) with or without PxP motif fusion peptide derived from adaptor proteins Mcp1, Ypt35 or Spo71.

Recombinant Rab8a expression and purification protocol as used by the Leschziner and Reck-Peterson Labs.

A method to monitor the recruitment of purified LRRK2 kinase onto planar lipid bilayers containing lipid-anchored Rab10 protein using Total Internal Reflection Fluorescence (TIRF) Microscopy.

Protocol for generating midbrain organoids from human iPSCs. In this protocol we describe the differentiation of human induced pluripotent stem cells (hiPSCs) into human midbrain-like organoids (hMLOs). This protocol has been developed based from several published protocols.

This protocol details Membrane Tube Image Analysis.

This protocol details about the Membrane Tube Assay.

MBP Pulldown Assay of ATG9A Truncations

This assay uses TMR labeled LRRK2 or LRRK1 RCKW to measure binding to microtubules in vitro

Assay to determine LRRK2 protein binding to microtubules.

Protocol for immunofluorescent staining for LRRK2 in cultured cells using the MJFF2 (c41-2) antibody.

We developed a method for assessing mitochondrial clearance in primary hippocampal neurons.

A protocol for rapidly purifying Golgi across various cell lines and tissues without any modification. The Golgi purified using this method are highly enriched, intact, contaminant-free and, depending on solubilisation buffer, could be used for various downstream applications, such as proteomics and metabolomics. Access the interview with Rotimi Fasimoye, PhD, about this protocol.

This protocol details about intracellular cytokine (ICS) staining.

Protocol for expressing LRRK1 and LRRK2 in insect cells.

This protocol describes the in vivo electrophysiology method for recording neuronal activity in mice.

This protocol describes immunohistochemical staining of fixed brain sections.

Imaris intensity-based protocol for Colocalization analysis.

Image quantification.