A fluorescence-based in vitro scrambling assay for yeast MCP1 and human XK
By Blythe Lloyd onDetailed procedure of purification, reconstitution, and scrambling assay for both MCP1 and XK.
Western blot protocol for detecting ATP10B in mouse/rat brain
By Michelle onThis protocol describes the procedure for detection of ATP10B in rat and mouse brain tissue by Western blotting
HyDrop-RNA v1.0
By Michelle onStep-by-step protocol for performing HyDrop-RNA. The duration of each step assumes an experienced protocol user. For a first-time user, we recommend doubling the expected time for each step.
Microfluidic Chip Production v1.1 V.2
By Michelle onProtocol for producing microfluidic chips used in HyDrop experiment.
Protocol for hippocampal neuronal cultures
By Blythe Lloyd onThis protocol details the procedure for preparation of neuronal cultures from mice hippocampi as it was performed in https://doi.org/10.1083/jcb.202010004 but can also be used to prepare cultures of cortical neurons.
Optogenetic experiments with iLID system
By Michelle onThis protocol details experiments with the iLID optogenetic system as performed to acutely recruit Miro to mitochondria in https://doi.org/10.1083/jcb.202010004.
Protocol for Image processing and analysis of VPS13D recruitment to mitochondria
By Michelle onThis protocol details the image processing and analysis of VPS13D recruitment to mitochondria as it was performed in https://doi.org/10.1083/jcb.202010004. The first part details the analysis of acute optogenetic recruitment, and the second part of basal recruitment under different conditions.
Hippocampal Neuronal Culture
By Blythe Lloyd onThis protocol describes the procedure for hippocampal neuronal cultures from new - born mouse pups.
Microscopy-based bead protein-protein interaction assay
By taliag onAn assay to study protein-protein interaction
Assessing enrichment of proteins in the mitochondrial fraction in HEK cells
By taliag onThis is a method for measuring protein enrichment on mitochondria in various conditions. In the resulting Western blot, one can assess the level of contamination of other organelles in the enrichment prep.
HyDrop Bead Generation & PCR Barcoding v1.0
By Blythe Lloyd onProtocol for producing dissolvable barcoded hydrogel beads used in HyDrop experiments.
TBK1 knockdown and rescue in Hela-M cells
By Blythe Lloyd onThis protocol transiently depletes TBK1 from HeLa cells and re-introduces a tagged TBK1 along with other relevant components of mitophagy. Because TBK1 is integral in many cellular processes, cells depleted of TBK1 exhibit poor health. Our protocol accomplishes ~70% depletion of endogenous TBK1 within 24 hours without excessive cell death.
VPS13D DNA plasmid generation
By Blythe Lloyd onThis protocol describes the basic molecular cloning technique utilized for the generation of VPS13D constructs in VPS13D bridges the ER to mitochondria and peroxisomes via Miro. This protocol and the enzymes included in it are commercialized by Takara Bio.