Maintenance of the recording chamber for non-human primate (Chamber cleaning and dura scraping)
By onThis protocol explains two procedures to maintain the recording chamber in nonhuman primates with craniotomies:(1) chamber cleaning and (2) dura scraping.
Stereotactic Viral Injection into Subthalamic Nucleus in Mice
By onThis protocol describes the surgical procedure of injecting AAV viruses into the subthalamic nucleus of a mouse. It contains information regarding the stereotactic setup, surgery preparation, anesthesia, craniotomy, injection, and recovery.
Handling and behavioral training protocol for in vivo electrophysiological and optogenetic manipulation of basal ganglia neurons in awake head-fixed mice
By onThis protocol details the habituation of animals and behavioral training of animals for in vivo electrophysiology or optogenetic experiments.
Fluorescent Immunolabelling for endogenous mouse LRRK2
By onThis protocol is used to label endogenous LRRK2 in mouse. It has been edited from West et al., 2014. Protocol optimized for fresh fixed brain tissue sectioned at 40 um.
Surface protein biotinylation
By onThis protocol describes surface protein labeling with biotin using EZ-Link Sulfo-NHSLC-Biotin. This chemical reacts with primary amines such as lysine but does not permeate cell membranes because of the charge. Thus, it only biotinylates surface proteins.
RNA isolation and qRT-PCR
By onThis protocol describes the RNA isolation from cultured cells and quantitative RT PCR.
Radiolabeled polyamine uptake in cells
By onThis protocol provides a technique to determine the radiolabeled polyamine uptake capacity in cells, via the acquisition of disintegrations per minute (DPM) using a Liquid Scintillation Counter.
Fluorescently labeled polyamine uptake (via Flow Cytometry)
By onAssess polyamine uptake capacities of a specific cell line after incubation with fluorescently labeled polyamines and mean fluorescence intensity acquisition via flow cytometry.
Microscopy-based bead protein-protein interaction assay.
By onProtocol for microscopy-based bead protein-protein interaction assay using GST- or mCherry-tagged baits and fluorescently-tagged preys. Requires purified proteins, monitors interactions in equilibrium, and allows quantification of fluorescent signal.
Immunohistochemistry of rTg4510 mouse brain
By onThe AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds.
Complex I activity assay
By onThis protocol is part of a Collection of protocols (dx.doi.org/10.17504/protocols.io.8epv593dng1b/v1) for the paper "Glucocerebrosidase, a Parkinson´s disease-associated protein, is imported into mitochondria and regulates complex I assembly and function" (https://doi.org/10.21203/rs.3.rs-1521848/v1)
In Vitro FSCV Testing of Carbon Fiber Electrodes to Characterize Functional Operation in Dopamine Detection
By onMethods to measure the performance characteristics of carbon fiber electrodes for neurochemical recording.
Co-immunoprecipitation protocol to study LRRK2 binding to Rab12 in a cell-based assay
By onThe immunoprecipitation of FLAG-tagged LRRK2 from whole cell lysates to assess its interaction with Rab12.
Characterization of Splay Characteristics of Carbon Fiber Electrode Threads (CFETs) in Agar Brain Phantom
By onMethods to measure deflection characteristics of carbon fiber electrodes in agar brain phantoms.
Lentivirus production for primary neuron transduction
By onThis protocol is associated with the following preprint, published on February 19th 2022: The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds Itika Saha, Patricia Yuste-Checa, Miguel Da Silva Padilha, Qiang Guo, Roman Körner, Hauke Holthusen, Victoria A. Trinkaus, Irina Dudanova, Rubén Fernández-Busnadiego, Wolfgang Baumeister, David W. Sanders, Saurabh Gautam, Marc I. Diamond, F. Ulrich Hartl, Mark S. Hipp bioRxiv 2022.02.18.481043; doi: https://doi.org/10.1101/2022.02.18.481043
DQ-BSA assay
By onThis protocol describes the DQ BSA assay for measuring the lysosomal proteolytic activity.
Live-imaging of axonal cargoes in human iPSC-derived neurons or mouse primary neurons
By onThe authors describe the procedure and equipment used for live imaging of axonal cargoes.
