EMG recordings-Rodrigues-Vaz and Athalye et al, 2025
By onThis is the standard protocol for recordings of bulk EMG used for Rodrigues-Vaz and Athalye et al, 2025.
Isometric forelimb task behavioral training
By onThis task is a self-paced task in which mice are reinforced for using their forelimb to apply force to an immobile joystick exceeding a threshold of duration and force.
Immunoprecipitation of NAP1-GFP
By onThis protocol describes the procedure to perform immunoprecipitation of NAP1-GFP from HAP1 cells.
GCase activity assay
By onProtocol measures GCase activity using 4-MUG substrate. GCase protein from a cellular model is diluted in acidic buffer to mimic lysosome pH. Adding lipid/detergent maintains enzyme activity. Taurocholate bile salt is included in the protocol.
Immunofluorescencent Staining of A-synuclein, TFEB, GCase and LAMP1 in treated cultured cells
By onProtocol used to detect a-synuclein, TFEB, GCase, and LAMP1 via immunofluorescence in SH-SY5Y cells with mutant SNCA gene.
Protocol — DQ™ Red BSA Lysosomal Activity Assay HCS Analysis
By onLysosomal proteolysis can be measured using self-quenched DQ™ Red BSA, followed by confocal imaging for spot-based quantification.
Protocol — Quantitative Real-Time PCR (qRT-PCR): 3K-SNCA cells
By onGene expression in 3K-SNCA SH-SY5Y cells was analyzed using Qiagen RNA extraction and TaqMan qRT-PCR on QuantStudio 1 platform for differentiated and undifferentiated cells.
Protocol — LysoTracker® Red DND-99 Live-Cell Staining 26 HCS Analysis
By onText summary: Live-cell lysosomal staining done with LysoTracker Red DND-99, followed by high-content confocal imaging using Opera Phenix Plus, and automated quantification in Harmony/Columbus.
Processing Serum, Plasma, Plasma EDTA from Whole Blood
By onProtocols cover serum, plasma, and plasma EDTA processing from whole blood samples.
Isolation of Classical Monocytes from PBMCs (MB Kit No. 130-117-337)
By onProtocol isolates classical monocytes from peripheral blood mononuclear cells.
PBMCs isolation with CT Tubes (8mL Whole blood)
By onProtocol isolates peripheral blood mononuclear cells using CPT tubes.
Dephosphorylation of phosphorylated Rab GTPases by PPM phosphatases
By onProtocol measures phosphoRab GTPase dephosphorylation with PPM phosphatases using purified phosphoRab GTPases. Substrates generated by phosphorylating Rab GTPases with MST3 kinase are included.
Flowcytometry analysis of lysosomal pulldown with anti-TMEM192 magnetic beads from PBMCs
By onMethod described for staining intact lysosomes with lysotracker for flow cytometry. Analysis provides quantitative indication of organelle enrichment in immunoprecipitation, serving as quality control.
GCase assay – Resorufin β-D-glucopyranoside
By onMethod presented for assessing lysosomal enzyme GBA1 activity using Resorufin substrate in low pH conditions to minimize non-lysosomal activity. Validated in GBA1 KO cells, compatible with various lysates and adjustable for different GBA1 levels.
p-S65-Ub sandwich ELISA
By onAssay measures Ubiquitin phosphorylation at Serine-65 in vivo using ELISA. Method previously detailed by Watzlawik et al. in their publication.
Cytokine Array
By onThe EMD MILLIPLEX‱ MAP Human Cytokine/Chemokine/Growth Factor Pane A – Immunology Multiplex Assay (Merck Millipore) is employed to simultaneously analyse TNF-α, IL-1β, IL 6, and INFγ with Bead-Based Multiplex Assay using the Luminex‱ technology.
Intracellular ROS measurement in iPSC-derived microglia
By onIntracellular ROS measurement in iPSC-derived microglia
