SNP Genotyping and ApoE Genotyping
By Michelle onThis protocol details the steps for DNA extraction from a human blood sample, quality control, and SNP and APOE genotyping. The protocol has been adapted from the PRoBaND SNP Genotyping and ApoE Genotyping Protocol
In situ architecture of the lipid transport protein VPS13C at ER-lysosomes membrane contacts
By Blythe Lloyd onPublication: Loss-of-function mutations in VPS13C are responsible for rare cases of familial early onset Parkinson’s disease. Using cryo-ET, the authors provide in-situ evidence for a bridge-model of VPS13 in lipid transport. View the original preprint.
DOPA pheomelanin is increased in nigral neuromelanin of Parkinson’s disease
By Blythe Lloyd onPublished: Neuromelanin in the substantia nigra may be a key factor contributing to dopaminergic neuron vulnerability in Parkinson’s disease. Neuromelanin consists of pheomelanin and eumelanin moieties. Here, authors investigated the relative composition and specific roles of pheomelanin and eumelanin moieties of NM in PD. View original preprint.
Staining of cells with GolgiTracker for Golgi flow cytometry analysis
By Michelle onMethod that allows the staining of intact Golgi using GolgiTracker for subsequent flow cytometry analysis. The analysis can provide a quantitative indication of the level of enrichment of the organelle immunoprecipitationand can be used as a quality control check for the purity of the immunoprecipitated Golgi.
Standard Operating Procedure: Vibratomes
By Michelle onThis Standard Operating Procedure (SOP) describes the safe use of vibratomes.
Statistical analysis
By Michelle onThis protocol describes the statistical analysis applied to the quantifications in Chang et al. 2021 SA paper.
Striatal dopamine measurement through HPLC
By Michelle onProtocol for striatal dopamine measurement in mouse brain
Subtomogram averaging and classification
By Michelle onThis protocol describes the procedure of subtomogram averaging and 3D classification of VPS13C rod-shaped densities inside cryotomograms of mammalian cells. Sub-tomogram averaging package i3 was used for 3D alignment and classification.
Surface protein biotinylation
By Michelle 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.
Unilateral intranigral AAV alpha synuclein mouse model
By Michelle onThis protocol details methods for the unilateral intranigral AAV alpha-synuclein injection.
Western blotting for LRRK2 signalling in macrophages
By Michelle onThis protocol describes the immunoblotting for components of the LRRK2 signalling pathway (LRRK2, LRRK2 pS935 and phospho-Rabs) using Invitrogen NuPage SDS-PAGE reagents and the BioRad Turbo Blot transfer system.
Yeast cells live fluorescence imaging
By Michelle onProtocol for sample preparation and live fluorescence imaging of yeast cells.
CoxII degradation assay to assess mitophagy
By Blythe Lloyd onThis protocol details the procedure to assess mitophagy by analyzing COXII degradation via Western blotting.