Protocol Archive

Traditional in vitro pulldown.

Pulse-chaseable method for quantifying autophagy flux.

Phosphorylation of Rab7 at S72 using TBK1.

Method of quantifying in vitro binding.

Affinity purification followed by size exclusion chromatography.

Autophagosome live cell imaging for quantification of PINK-PARKIN mitophagy.

Plasmid construction via restriction enzyme digest and ligation.

Traditional Western blotting technique.

Growth in DMEM supplemented with FBS and antibiotics.

The authors describe their method to isolate peripheral blood mononuclear cells (PBMCs) and perform rapid isolation of intact, tag-less lysosomes from PBMC homogenates using ball-bearing cell breakers and anti-TMEM192 -antibody coupled to magnetic beads.

This protocol can be used to immunoprecipitate lysosomes from commonly cultured cells such as mouse embryonic fibroblast, HEK293, and A549 cells expressing the LysoTag.

Methods detailing the recording setup using both electrophysiological (Ephys) and fast-scan cyclic voltammetry (FSCV) are described.

This protocol describes how the authors prepare human iPSC-derived cells, that have been differentiated into cortical neuronal progenitors, for transplantation into the brain of immunocompromised athymic mice. Neuronal progenitors mature within the mouse brain and are used to study the pathogenesis of Parkinson’s disease.

This protocol details the primary neuron culture procedures from embryonic and postnatal mice.

These procedures are done in the context of calcium imaging experiments in non-human primates. The authors inject a solution containing AAV-GCaMP6 into the supplementary motor area (SMA) of the animal to be tested and then insert the GRIN lens into the injected area during the same session.

This protocol details ventral midbrain genomic PCR.

This protocol details the staining for free floating mouse brain sections.

This protocol details the immunofluorescence staining in mouse brain tissue sections.

Protocol for detecting pRab proteins in mouse brain tissue.

Protocol for performing ImmPRESS immunostaining with alkaline phosphatase labeling after BaseScope In Situ Hybridization mouse brain sections.