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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
Rab8a expression and purification
Recombinant Rab8a expression and purification protocol as used by the Leschziner and Reck-Peterson Labs.
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LRRK2 RCKW protein purification
Protein purification protocol for tag-less LRRK2RCKW as done by Leschziner and Reck-Peterson Labs. Same protocol can be used to purify LRRK1RCKW as well. Original protocol by David Snead. Modified by Yu Xuan Lin and Mariusz Matyszewski for publication.
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LRRK1 expression and purification
Protein purification protocol for full-length LRRK1 as done by Leschziner and Reck-Peterson Labs.
Original protocol by Janice M. Reimer and Yu Xuan Lin.
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Insect Cell Protocol for LRRK1 and LRRK2 Expression
Protocol for expressing LRRK1 and LRRK2 in insect cells.
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LRRK2 microtubule sedimentation binding assay
Assay to determine LRRK2 protein binding to microtubules.
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LRRK2RCKW Widefield fluorescence microtubule binding assay
This assay uses TMR labeled LRRK2 or LRRK1 RCKW to measure binding to microtubules in vitro
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Preparation of LRRK2 RCKW cryo-EM grids
This is Leschziner’s Lab updated protocol for making cryo-EM grids for LRRK2 RCKW. This protocol, when using lower protein concentration, results in better monomer and dimer formation than the old protocol.
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Preparation of LRRK1 RCKW cryo-EM grids
Protocol used to create LRRK1 RCKW grids for cryo-EM used in Snead, Matyszewski, Dickey et al.
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Quick guide to use paceTOMO for cryo-ET data collection from Titan Krios
This quick guide provides key minimal steps for preparing the Titan/SerialEM for the tomogram data collection on lamella or in vitro specimens with a K3 camera. paceTOMO routine is also included for a typical tomogram data collection session. Please note that this is not an exhaustive guide, but summarizes the order of key steps.
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Image processing of full-length monomeric LRRK2
This protocol assumes that 2000 to 4000 movies of full-length LRRK2 embedded on vitreous ice are collected with an electron microscope equipped with a direct detector. It focuses primarily on the monomeric population of LRRK2 and has been leading to 3-4 Å resolution structures.
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