Sample preparation for aCGH karyotyping
By onThis protocol describes the standard procedure preparing cell pellets from human pluripotent stem cells (hPSCs) cultured on MEFs for outsourced aCGH karyotyping.
iPSC differentiation into Macrophages
By onThis protocol describes iPSC differentiation into macrophages.
Mechanisms Controlling Selective Elimination of Damaged Lysosomes
By onLysophagy, triggered by membrane rupture, involves galectins binding to lysosomal contents to promote autophagic recycling. Damaged lysosomes are ubiquitylated, leading to autophagosome formation for degradation in healthy lysosomes.
Vesicular dysfunction and pathways to neurodegeneration
By onIn this review, the pathways that have emerged as critical for neuronal survival in the human brain are discussed, illustrating the diversity of proteins and cellular events with three molecular case studies from different neurological diseases.
pBMN_HA-TBK1 (E696K)
By onPlasmid: Used for the expression of TBK1 carrying E696K mutation (SMC Internal No. 1998).
pBMN_HA-TBK1 (kinase dead – D135N)
By onPlasmid: Used for the expression of TBK1 carrying a kinase dead mutation D135N (SMC Internal No. 1997).
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.
In Vivo Electrophysiology (Mouse)
By onThis protocol describes the in vivo electrophysiology method for recording neuronal activity in mice.
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.
Immunohistochemistry
By onThis protocol describes immunohistochemical staining of fixed brain sections.
pCAG-MBP-ATG9 (1-830)
By onPlasmid: Expresses human ATG9 C-terminal truncation mutant in mammalian cells.
Culture and transfection of HEK293T cells
By onThis protocol describes a standard procedure culturing and transfecting HEK293T cells Protocol overview: A. Culturing HEK293T cells B. Transfection of HEK293T cells with Lipofectamine 2000
Staining of cells with GolgiTracker for Golgi flow cytometry analysis
By onMethod that allows the staining of intact Golgi using GolgiTracker for subsequent flow cytometry analysis.
Px330-EGFP-LRRK2-CRISPR/Cas9
By onIt can be used to introduce LRRK2-G2019S mutation using CRISPR/Cas9 based genome editing.
Highly efficient generation of isogenic pluripotent stem cell models using prime editing
By onPrime editing (PE) simplifies creating human pluripotent stem cell (hPSC) disease models by optimizing mRNA delivery with editing efficiency increased up to 13-fold, enabling correction or introduction of Parkinson's disease mutations in hPSCs.
pGST2-GST-TEV-OPTN S177D S473D
By onPlasmid for the expression and purification of GST-TEV-OPTN S177D S473D. Internal Ref: SMC1588
Single cell analysis of iPSC-derived midbrain organoids
By onThe following script was used for analysis of gene corrected (GC) versus GBA1 mutant (MUT) midbrain organoids. The purpose was to combine, filter, integrate, and identify clusters and differentially expressed genes sets. This is part of a Collection of protocols (dx.doi.org/10.17504/protocols.io.8epv593dng1b/v1) for the paper "Glucocerebrosidase, a Parkinson´s disease-associated protein, is imported into mitochondria and regulates complex I assembly and function" (https://doi.org/10.21203/rs.3.rs-1521848/v1)
