Electron microscope sample preparation technique_V2
By Blythe Lloyd onThere are eight main steps in preparing EM sample, including fixation, en bloc staining, agarose embedding, dehydration, infiltration, resin embedding, resin-embedded sample trimming and ultrathin sectioning. This EM sample preparation protocol is designed for processing cultured cell sample.
Design and preparation of synthetic reference peptides for APP/Aβ TOMAHAQ proteomics, version 2
By Blythe Lloyd onThis protocol describes the design and preparation of synthetic reference peptides for APP/Aβ TOMAHAQ proteomics.
Deep Brain Stimulation (DBS) Implant
By Blythe Lloyd onThis protocol describes the steps for making implants used for deep brain stimulation (DBS) in rodents.
Crystallization, data collection and structure solution of PxP-VAB
By Blythe Lloyd onProtocol lists the steps to obtain PxP(Mcp1ct)-VABct crystals, X-ray diffraction data acquisition and obtaining the structure solution using molecular replacement with a model generated by AlphaFold2.
Sample Collection and Measurement of Serum Neurofilament Light (NfL)
By Michelle onThis protocol details the steps for the preparation of serum from human blood samples and the measurement of NfL concentration using the NF-Light Advantage kit on the HD-X Analyzer (Quanterix, Billerica, MA).
Single-cell dissociation of brain organoids
By Michelle onSingle-cell dissociation of brain organoids
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
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.