pCAG- WIPI2dC93E-cs- TEV-STREP
By Michelle onMammalian expression of human WIPI2d C93E with C-terminal Strep.
pCAG- WIPI2dH85E-cs-TEV-STREP
By Michelle onMammalian expression of human WIPI2d H85E with C-terminal Strep.
Collection of protocols for paper: “Glucocerebrosidase, a Parkinson´s disease-associated protein, is imported into mitochondria and regulates complex I assembly and function”
By Blythe Lloyd onThis is a collection of protocols used in a recent pre-print by the Deleidi Lab, Team Schapira. You can access pre-print at https://doi.org/10.21203/rs.3.rs-1521848/v1
pCAG- WIPI2dI92E-cs- TEV -STREP
By Michelle onMammalian expression of human WIPI2d I92E with C-terminal Strep.
pCAG- WIPI2dK128E- cs-TEV -STREP
By Michelle onMammalian expression of human WIPI2d K128E with C-terminal Strep.
Cell line construction and maintenance for Lyso-IP and Endo-IP analysis of amyloid precursor protein processing
By Blythe Lloyd onLyso-IP is a method that allows for the isolation of lysosomes for proteomics and metabolomics (dx.doi.org/10.17504/protocols.io.bybjpskn; dx.doi.org/10.17504/protocols.io.bx9hpr36). We have developed an analogous approach for purification of early/sorting endosomes (Endo-IP).
pCAG- WIPI2dK88E-cs- TEV-STREP
By Michelle onMammalian expression of human WIPI2d K88E with C-terminal Strep.
pCAG- WIPI2dR108E- cs-TEV -STREP
By Michelle onMammalian expression of human WIPI2d R108E with C-terminal Strep
pCAG- WIPI2dR125E- cs-TEV -STREP
By Michelle onMammalian expression of human WIPI2d R125E with C-terminal Strep
3D-correlative FIB-milling and Cryo-ET of Autophagic structures in Yeast Cells
By Michelle onThis protocol describes how to plunge-freeze yeast on EM grids and how to target autophagic structures by combining cryo confocal fluorescence data to FIB-milling and tomogram acquisition.
Cell culture on EM grids and fluorescence microscopy imaging
By Blythe Lloyd onThis protocol describes the general procedure of seeding mammalian cells on EM grids and confocal fluorescence microscopy imaging of grids for subsequent cryo-tomography experiments, performed.
Binding of Rab29 to the LRRK2 Armadillo domain by Microscale Thermophoresis
By Blythe Lloyd onMicroscale thermophoresis (MST) is a powerful tool to measure the affinities of interactions between proteins. We present here our method for determining the binding of Rab29 GTPase to the LRRK2 (Leucine rich repeat kinase 2) N-terminal Armadillo domain.
Behavioral Testing – Open Field and Dyskinesia Scoring
By Blythe Lloyd onThis protocol describes behavioral testing to assess motor deficits in mice. It includes open field locomotor testing and scoring of levodopa-induced dyskinesia.