Purification of the PE2 nCas9-RT protein
By Michelle onThis protocol describes the process of expressing and purifying the nicking Cas9-MMLV RT fusion protein for prime editing.
Purifying Vps13-VAB domain proteins
By Michelle onIn 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.
Rab8a expression and purification
By Michelle onRecombinant Rab8a expression and purification protocol as used by the Leschziner and Reck-Peterson Labs.
Reconstitution of LRRK2 membrane recruitment onto planar lipid bilayers
By Michelle onA 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.
Midbrain-like Organoids generation from hiPSCs
By Blythe Lloyd onProtocol 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.
LRRK2RCKW Widefield fluorescence microtubule binding assay
By Blythe Lloyd onThis assay uses TMR labeled LRRK2 or LRRK1 RCKW to measure binding to microtubules in vitro
LRRK2 microtubule sedimentation binding assay
By Blythe Lloyd onAssay to determine LRRK2 protein binding to microtubules.
LRRK2 Immunofluorescent staining
By Blythe Lloyd onProtocol for immunofluorescent staining for LRRK2 in cultured cells using the MJFF2 (c41-2) antibody.
Investigation_of_mitophagy_in_Hippo_neurons
By Blythe Lloyd onWe developed a method for assessing mitochondrial clearance in primary hippocampal neurons.
Introducing GolgiTag to Cells and Immunoprecipitation of Golgi
By Blythe Lloyd onA 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.
Intracellular Cytokine (ICS) Staining Protocol
By Blythe Lloyd onThis protocol details about intracellular cytokine (ICS) staining.
Insect Cell Protocol for LRRK1 and LRRK2 Expression
By Blythe Lloyd onProtocol for expressing LRRK1 and LRRK2 in insect cells.
The emerging role of LRRK2 in tauopathies
By Blythe Lloyd onReview: Authors review the emerging evidence and discuss the potential impact of LRRK2 dysfunction on tau aggregation, lysosomal function, and endocytosis and exocytosis.
Pathological structural conversion of α-synuclein at the mitochondria induces neuronal toxicity
By Blythe Lloyd onPublished: Using a single molecule FRET sensor, the authors track the intracelleular conformational states of alpha-synucelin and where these occur in the cell. The study suggests that oligomerization happens at the mitochonodria triggering neuronal toxicity. View original preprint.
In Vivo Electrophysiology (Mouse)
By Blythe Lloyd onThis protocol describes the in vivo electrophysiology method for recording neuronal activity in mice.
Immunohistochemistry
By Blythe Lloyd onThis protocol describes immunohistochemical staining of fixed brain sections.
Imaris Coloc Protocol
By Blythe Lloyd onImaris intensity-based protocol for Colocalization analysis.
