Microscopy-based GSH bead protein-protein interaction assay
By onProtocol describing the Microscopy-based GSH bead protein-protein interaction assay
Cellular lipid uptake with flow cytometry readout
By onCellular lipid uptake with flow cytometry readout
GST Bead pulldown Assay
By onGST Pulldown Assay for recruitment of bait proteins to GST labeled prey proteins. Prey proteins can be purified or from lysate.
Endosomal and lysosomal immunoprecipitation for proteomics, lipidomics, and TEM
By onHere, authors describe an approach for purification of early/sorting endosomes, providing a means by which to examine early aspects of the endolysosomal system (Endo-IP) and to combine this with lysosome purification using Lyso-IP.
Immunostaining of iPSC-derived neurons for quantification of synaptic proteins
By onHere, the authors fix, permeabilize, and stain human iPSC-derived neurons for the purpose of observing and quantifying somal proteins of interest.
Isolation and Processing of Embryonic and Postnatal Brains
By onThis protocol describes: -Harvesting embryos from females from embryonic day 10 to embryonic day 18 -Brain isolation of embryonic (e10-e18) and postnatal brains (p0-p30) -Processing and Freezing of embryonic and postnatal brains
Sectioning of Mouse Brain by Microtome
By onThis protocol describes how to use the microtome to prepare and slice mouse brain sections for Immunohistochemistry.
Quantifying Acetylcholinesterase activity in striatal brain tissue with DTNB assay
By onThis protocol describes the steps to measure acetylcholinesterase (AChE) activity in mouse striatal brain tissue, including how to store tissue samples, extract AChE and obtain Michaelis-Menten like plots to measure the rate of AChE activity.
Midbrain viral injections for striatal fiber photometry in mice
By onThis protocol describes how to: - Inject AAV viruses into the midbrain (SNc: substantia nigra pars compacta, or VTA: ventral tegmental area) of mice - Train mice for head-fixed running on a cylindrical treadmill, and to receive rewards and air puffs while head-fixed - Implant head plates on mice for head-fixation during behavior (optional) While this protocol is focused on injections of AAV viruses into the midbrain, it can be easily modified to inject into other brain regions or other viruses.
Mitochondrial complex activity assays
By onMitochondria complex activity assays measure the activity levels of the different complexes of the mitochondrial electron transport chain (ETC).
Immunoblotting of macrophages and microglia
By onThis protocol describes the preparation from cell lysate from cultured cells and immunoblotting procedure.
mCherry-YIPF4 Immunoprecipitation V2
By onThe protocol outlines how to immunoprecipitate mCherry-YIPF4 for further analysis.
Endogenous coimmunoprecipitation
By onProtocol used for immunoprecipitation of HSP60 and LONP1 in HEK cells to show the interaction with V5-Flag-tagged WT-GCase
Gene editing of YIPF4, CALCOCO1, and ATG7 in HeLa cells
By onThe text outlines the process of generating YIPF4, FIP200, and ATG7 knockout cell lines in HeLa cells through CRISPR-Cas9 technology.
Transferrin uptake assay to measure Clathrin-mediated endocytosis in HIPCS derived neurons
By onThis protocol is used to measure Transferrin uptake as a way to investigate Clathrin-mediated endocytosis in HIPCS derived neurons, or other cell types.
In Vivo Carbon Fiber Electrode Thread (CFET) Implantation and Testing Procedures
By onProcedures for implanting and testing CFET devices in rats are described.
Primary neuron culture for live imaging of axonal cargoes
By onThis protocol describes the preparation and culture of mouse primary cortical neurons for live-imaging experiments. Neurons were transfected 16-24 hours before imaging using Lipofectamine 2000.
Microscopy-based evaluation of mtKeima flux in hESC-derived Ctrl and FBXO7-/- iNeurons
By onProtocol for the microscopy-based evaluation of mtKeima flux in hESC-derived Ctrl and FBXO7-/- iNeurons PARK15/FBXO7 is dispensable for PINK1/Parkin mitophagy in iNeurons and HeLa cell systems: https://www.embopress.org/doi/full/10.15252/embr.202256399
