Glucose Tolerance Test
By onThe glucose tolerance test is performed to determine how quickly the glucose is cleared from the blood. It is used to test for diabetes or insulin resistance.
Striatal Cannula Window Implantation
By onThis protocol describes how to implant a cannula window over the dorsolateral striatum in mice to enable two-photon imaging of dopamine axons in the dorsal striatum.
Workflow for proteomic analysis of purified lysosomes in cells lacking GRN V.3
By onAssociated with the following preprint (published September 30th 2021): Progranulin deficiency results in reduced bis(monoacylglycero)phosphate (BMP) levels and gangliosidosis Sebastian Boland, Sharan Swarup, Yohannes A. Ambaw, Ruth C. Richards, Alexander W. Fischer, Shubham Singh, Geetika Aggarwal, Salvatore Spina, Alissa L. Nana, Lea T. Grinberg, William W. Seeley, Michal A. Surma, Christian Klose, Joao A. Paulo, Andrew D. Nguyen, J. Wade Harper, Tobias C. Walther, Robert V. Farese Jr. bioRxiv 2021.09.30.461806; doi: https://doi.org/10.1101/2021.09.30.461806
In vitro kinase assay: phosphorylation of PI3Kc1 by TBK1and ULK1
By onProtocol describing in vitro kinase assay, in which PI3Kc is tested for phosphorylation by TBK1 and ULK1 complex either by western blotting (S29 on ATG14L subunit) or mass spectrometry analysis.
Brain processing, slicing and immunohistochemistry protocol
By onThis protocol is a step by step procedure from collecting brain samples to immunohistochemical staining and mounting brain slices.
Pole Test to assess motor coordination in parkinsonian mice
By onThe pole test evaluates the motor coordination ability of a mouse by having them grasp a pole and then descend to its home cage.
Mouse Ovariectomy
By onThis protocol describes the steps to perform an ovariectomy in a mouse. This procedure is used to induce menopause.
HyDrop Bead Generation & PCR Barcoding v1.0
By onProtocol for producing dissolvable barcoded hydrogel beads used in HyDrop experiments.
Mitochondrial Antigen Presentation in RAW macrophages
By onThis protocol details methods for 3-day Mitochondrial Antigen Presentation Assay in a murine macrophage cell line (RAW) that expresses a glycoprotein B (gB) of herpes simplex virus 1 (HSV1) targeted to the mitochondrial matrix (mito-gB). A gB-specific CD8+ T cell hybridoma recognizing the gB498–505peptide loaded on MHC class I molecules is also used to monitor antigen presentation through a beta-galactosidase assay kit.
LC3-lipidation-assay
By onProtocol for an in vitro LC3 lipidation assay using purified proteins and synthetic liposomes.
Measurement of GLP-1 release in cell supernatant from Hutu-80 enteroendocrine cells via ELISA
By onMeasurement of GLP-1 release in cell supernatant from Hutu-80 enteroendocrine cells via ELISA
Creating pooled CRISPR-Cas9 knock-outs in NIH-3T3 cells
By onTo validate a genome wide CRISPR screen, the authors select the top hits and create lentiviruses to validate the hits.
Western blot – alpha-synuclein
By onThis protocol describes how to detect alpha-synuclein in protein derived from STC-1 cells by western blot using DAB/peroxidase or ECL to visualize the bands.
Mouse Brain Tissue Collection and Analysis
By onThis protocol describes the dissection and collection of coronal sections of the striatum and midbrain from a mouse brain. The tissue can be used in a number of applications and here we describe two: the measurement catecholamine levels using high-performance liquid chromatography (aided by a neurochemistry core) and the distribution of the vesicular monoamine transporter (VMAT2) using a radioligand binding assay.
Challenging Beam Test
By onThe challenging beam test is used to measure motor performance, coordination, and balance.
Halo-LC3B processing assay to assess autophagy
By onThis protocol details Halo-LC3B processing assay to assess autophagy.
Wire Hang Test
By onThis behavioral test is designed to assess early motor deficits associated with muscle and grip strength.
Reconstitution of LRRK2 membrane recruitment onto planar lipid bilayers
By onA method to monitor the recruitment of purified LRRK2 kinase onto planar lipid bilayers containing lipid-anchored Rab10 protein using Total Internal Reflection Fluorescence (TIRF) Microscopy.
