Brain processing, slicing, and immunohistochemistry protocol
By Julia Julia Leonard onThis protocol is a step by step procedure from collecting brain samples to immunohistochemical staining and mounting brain slices.
Handling and behavioral training protocol for in vivo electrophysiological and optogenetic manipulation of basal ganglia neurons in awake head-fixed mice
By Julia Julia Leonard onThis protocol details the habituation of animals and behavioral training of animals for in vivo electrophysiology or optogenetic experiments.
Confocal imaging on Nikon AXR confocal microscope
By Julia Julia Leonard onThis protocol provides a step by step protocol to acquire images on Nikon AXR confocal set-up.
Immunofluorescence staining
By Julia Julia Leonard onThe protocol describes immunofluorescence staining on brain sections.
Automated immunohistochemistry staining
By Julia Julia Leonard onThe protocols describe the steps for automated immunohistochemistry staining using the Ventana Discovery® XT Immunostainer.
Luminescent conjugated oligothiophenes (LCO) staining
By Julia Julia Leonard onThe protocol describes Luminescent Conjugated Oligothiophenes staining on brain sections.
Protocol Collection: Combinatorial selective ER-phagy remodels the ER during neurogenesis
By Julia Julia Leonard onThis molecular inventory of ER proteome remodeling and versatile genetic toolkit provides a quantitative framework for understanding contributions of individual ER-phagy receptors for reshaping ER during cell state transitions.
Lentivirus preparation for neuronal transdifferentiation
By Julia Julia Leonard onThis is a protocol for the preparation of Lentiviruses.
Human primary fibroblast culture
By Julia Julia Leonard onThis is a protocol for human primary fibroblast culture.
Neuronal transdifferentiation from human primary adult fibroblasts
By Julia Julia Leonard onThis is a protocol for the direct conversion of human primary fibroblasts into neurons using a combination of transcription factor- and small molecule-based approach. The majority of converted neurons showing characteristics of cortical glutamatergic neurons.
Cytokine profiling analysis on conditioned medium of human neurons using Luminex multiplex assay
By Julia Julia Leonard onThis protocol is used to identify secreted inflammatory factors by cytokine profiling of the conditioned medium from human transdifferentiated neurons of healthy donors and AD patients.
Immunohistochemistry on free-floating and paraffin-embedded tissue sections
By Julia Julia Leonard onThis protocol is used for free-floating frozen (30-50 microns) and paraffin-embedded (10 microns) tissue sections.
Fast scan cyclic voltammetry
By Julia Julia Leonard onThis protocol describes the ex vivo fast-scan cyclic voltammetry (FSCV) technique for detecting dopamine in the dorsal striatum in mice.
Behavioral tests in rodents coupled with dopamine signaling manipulations
By Julia Julia Leonard onThese two protocols describe behavioral tests associated with Parkinson’s disease phenotypes in mice. The striatal motor learning protocol (rotarod test) is a two-phase paradigm assessing striatal motor learning under reversible dopamine antagonism.
Viral-mediated short-hairpin RNA knockdown
By Julia Julia Leonard onStereotaxic injection of viral vectors for gene knockdown experiments: The AAV vectors for knocking down the gene of interest were custom-made with Vector Biolabs using AAV-GFP-U6 vector for AAV1 packaging (Cat no, 7040, Vector Biolabs). They were then unilaterally injected into the striatum of mice as the protocol details.
Preparation of fresh paraformaldehyde for mouse perfusion and tissue fixation
By Julia Julia Leonard onPreparation of fresh paraformaldehyde for mouse perfusion and tissue fixation.
Unconventional secretion of alpha-synucein mediated by palmitoylated DNAJC5 oligomers
By Julia Julia Leonard onThis protocol, carried out according to the manufacturer’s instructions, uses a classical ELISA experimental design with sensitive electrochemical detection to provide a measure of 𝜶-synuclein concentration in the conditioned media of cultured iPSC-derived human cells.
Detection of accessible cholesterol in primary cilia using purified His-ALOD4-mNeon in 3T3 Fibroblasts
By Julia Julia Leonard onThe protocol described here is based upon previously established methods (Kinnebrew et al., 2019; Johnson and Radhakrishnan, 2021) and has also been used successfully to label cilia in RPE cells.
Preparation of soluble and insoluble mitochondrial protein fractions for mass spectrometry analysis
By Emma Sherrell onPreparation of soluble and insoluble mitochondrial protein fractions from HeLa cells for mass spectrometry analysis.
Transfection by electroporation of GFP-LRRK2 and Immunofluorescent imaging of MEFs VPS35 (D620N) mutants stably expressing LysoTag
By Emma Sherrell onThe authors investigated the colocalization of GFP-LRRK2 with a lysosomal localized TMEM192-3xHA.