Ubiquitin immunoprecipitation using an anti-ubiquitin nanobody
By onThis protocol describes a method to detect ubiquitination on a protein of interest. This technique relies on immunoprecipitation (IP) of ubiquitinated proteins from a cell lysate through the use of an anti-ubiquitin nanobody coupled to agarose beads. The eluate can be run on SDS-PAGE to determine whether the protein of interest was recovered in the IP and, therefore, ubiquitinated.
Immunofluorescence free-floating rat brain cryosections
By onProtocol for immunofluorescence on rat brain cryosections
Immunohistochemistry free-floating rat brain cryosections
By onProtocol for immunohistochemistry on rat brain cryosections.
Coupling of TMEM192 antibody to MyOne™ Epoxy Dynabeads™
By onThis protocol describes the coupling of rabbit monoclonal TMEM192 antibody to MyOne™ Epoxy Dynabeads™ . The coupled beads generated using this protocol can be used for the isolation of untagged lysosomes from cells and tissues.
Free floating immunofluorescence protocol on mouse brain sections for tau pathology
By onThis protocol describes the process of immunofluorescently labelling tau and phospho-tau in athymic mouse brains that have been grafted with human-disease derived iPSC's. This protocol can be adapted for other tissue types and antibody pairings.
Preparation and RNAscope-labeling of fresh mouse midbrain tissue
By onThe authors describe preparing and labeling coronal sections of mouse substantia nigra (SNc) and ventral tegmental area (VTA) for probes targeting the genes Slc17a6 (VGLUT2), Slc32a1 (VGAT), and Slc18a2 (VMAT2).
Preparation of Recording ACSF (artificial cerebrospinal fluid) for slice electrophysiology
By onPreparation of Recording ACSF Recording ACSF is used for incubating brain slices during cell-attached or whole-cell electrophysiological recordings of neurons.
Preparation of Holding ACSF (artificial cerebrospinal fluid) for slice electrophysiology
By onPreparation of Holding ACSF Holding ACSF is used for incubating brain slices prior to cell-attached or whole-cell electrophysiological recordings of neurons.
Isolation of lysosomes using the Tagless LysoIP method in PBMCs
By onDetailed protocol for rapid isolation of intact lysosomes (including from clinical samples) for subsequent -omics analyses.
Sample preparation protocol for proteomic analysis of isolated lysosomes and whole cell extracts
By onHere, the authors describe a step-wise protocol for proteomic sample preparation derived from cell line models or isolated human cells. The protocol has been optimized for organelle pulldown preps.
Mouse tissue collection
By onThis protocol is about collecting mouse tissues for immunohistochemistry or biochemistry experiments. While it is mainly used for brain tissues, it also can be used for other visceral tissues.
Mouse sample collection for metabolomics studies
By onProtocol used in the Mazmanian lab for collecting brain and gut tissues and plasma from mouse for metabolomics.
A method for RNA extraction, cDNA synthesis and and quantitative PCR from NIH-3T3 fibroblasts
By onProtocol for RNA extraction, cDNA synthesis, and qPCR from NIH-3T3 fibroblasts outlined. Method applicable to mouse embryonic fibroblasts as well.
DNA extraction protocol for saliva samples obtained from Omigene oral OM device using the Qiagen QiaAMP Powerfecal DNA kit for shotgun metagenomic sequencing
By onThis protocol describes the procedure for DNA extraction of saliva samples using the Omnigene OM device and the Qiagen QiaMP Powerfecal DNA PRO kit for shotgun metagenomic sequencing.
Lysosomal isolation protocol
By onProtocol for isolating lysosomes from cultured cells is described.
Immunofluorescent Labeling of Preformed Fibril Protein Protocol
By onProtocol outlines use of AlexaFluor 488 kit to label alpha synuclein preformed fibrils for fluorescence, facilitating future applications.
