CXCL1 supplementation on iPSC-Derived Midbrain Neurons
By onThe protocol studies the impact of different levels of recombinant human CXCL1 on midbrain neurons from iPSCs using conditioned media from ATP13A2WT astrocytes.
Fluorescently labeled polyamine uptake (via confocal microscopy)
By onMethod visualizes polyamine uptake in cells with fluorescently labeled polyamines & confocal microscopy.
Fluorescently labeled polyamine uptake (via flow cytometry)
By onSupplemental video in ASAP Protocol Particulars series provides extra context and tips from protocol authors for Aligning Science Across Parkinson's project.
Quantification of Cell Death and Cytotoxicity Using LDH-Glo™ Cytotoxicity Assay
By onThe LDH-Glo™ Cytotoxicity Assay quantifies cell death and cytotoxicity in cultured cells by measuring LDH released from damaged cells.
CoxII degradation assay to assess mitophagy
By onThis protocol details the procedure to assess mitophagy by analyzing COXII degradation via Western blotting.
Protocol for in vivo electrophysiological and optogenetic manipulation of basal ganglia neurons in awake head-fixed mice
By onStep by step procedure for in vivo electrophysiological and optogenetic manipulation of basal ganglia neurons in awake head-fixed mice.
Indirect Proximity Ligation Assay (PLA) – Brightfield
By onThe authors describe the PLA protocol that is routinely used in the laboratory to detect nitrated alpha-synuclein and nitration of mitochondrial enzymes such as SOD2 and the mitochondrial complex 1 subunit NDUFB8.
Binding of Rab29 to the LRRK2 Armadillo domain by Microscale Thermophoresis
By onMicroscale thermophoresis is a powerful tool to measure the affinities of interactions between proteins. We present here our method for determining the binding of Rab29 GTPase to the LRRK2 N-terminal Armadillo domain.
RNAScope in situ hybridization (ISH) multiplex fluorescent
By onInstructions to perform ISH using RNAscope kit from ACDBio company optimized for frozen mouse brain and nodose ganglia sections.
Single cell survival assay
By onThis protocol describes the experimental procedure used to measure single cell survival rates post nucleofection of human pluripotent stem cells (hPSCs). General Notes: 1. Throughout these protocols, the term hPSC is used to collectively refer to both hiPSCs and hESCs. All described procedures have been tested and work equally well for hiPSCs and hESCs.
Assaying starvation-induced autophagy in HeLa cells
By onA method for assaying starvation-induced autophagy in HeLa cells that have been transfected with Halo-tagged constructs.
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.
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.
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.
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.
