Native gel lipid binding assay
By onThis protocol details how to perform a native gel lipid binding assay to evaluate the number of lipids bound per molecule of 3xFLAG-SHIP164Δ901-1099
Co-immunoprecipitation using GFP-trap
By onProtocol describing the procedure to perform co-immunoprecipitation experiments in HEK293 cells using GFP-trap beads
Detection of Tau ubiquitylation
By onThis protocol is associated with the following preprint, published on February 19th 2022: The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds Itika Saha, Patricia Yuste-Checa, Miguel Da Silva Padilha, Qiang Guo, Roman Körner, Hauke Holthusen, Victoria A. Trinkaus, Irina Dudanova, Rubén Fernández-Busnadiego, Wolfgang Baumeister, David W. Sanders, Saurabh Gautam, Marc I. Diamond, F. Ulrich Hartl, Mark S. Hipp bioRxiv 2022.02.18.481043; doi: https://doi.org/10.1101/2022.02.18.481043
Rapalog-induced chemical dimerization experiments
By onThis protocol describes how to conduct Rapalog-induced chemical dimerization experiments.
RNA extraction and quantitative PCR to assay inflammatory gene expression
By onProtocol for RT-qPCR to test levels of NF-kB response genes in a cell model transiently over-expressing Parkin. We found upregulation of key pro-inflammatory genes normalized to housekeeping gene, Gapdh.
Open Field Test to assess motor coordination in a mouse parkinsonian model
By onThe Open Field task is a simple sensorimotor test used to determine general activity levels, gross locomotor activity, and exploration habits in rodent models of CNS disorders. Open Field Test
Coating coverslips for cell culture
By onCoated coverslips provide a nourishing adherent surface for cell culture. This protocol provides step by step instruction on how to coat coverslips for cell culture.
Purification and analysis of SKP1-FBXO7 complexes
By onProtocol for the biochemical purification and analysis of SKP1-FBXO7 complexes.
Confocal imaging on Nikon AXR confocal microscope
By onA step by step protocol to acquire images on Nikon AXR confocal set-up.
Endogenous tagging of the YIPF4 gene with mNEON Green and imaging of these cells by microscopy
By onProtocol for tagging YIPF4 gene with mNEON in cells using CRISPR. Method involves editing N-terminus of YIPF4 in WT and FIP200-/- HEK293T cells. Imaging mNEON-YIPF4 in Golgi with GOLGB1 marker is also detailed.
Microfluidic Chip Production v1.1 V.2
By onProtocol for producing microfluidic chips used in HyDrop experiment. Visit for more info
Mito and Glycolysis Stress Tests for Enteroendocrine Cells – Hutu-80
By onMito and glycolysis stress tests for enteroendocrine cells - Hutu-80
Headplate surgery protocol for in vivo electrophysiological and optogenetic manipulation of basal ganglia neurons in awake head-fixed mice
By onStep by step protocol for headplate surgery for in vivo electrophysiological and optogenetic manipulation of basal ganglia neurons in awake mice.
GFP Immunoprecipitation and Sample Preparation for Tandem Mass Tag (TMT) Mass Spectrometry Analysis
By onWe describe a method to identify potential interactors of any Green Fluorescent Protein (GFP) tagged protein expressed in mammalian cells by GFP immunoprecipitation coupled to Tandem Mass Tag (TMT) mass spectrometry analysis. As an example, we used a GFP-tagged phosphoRab interactor protein (RILPL1-GFP), and its non-binding mutant (RILPL1 -GFP, which cannot interact with phosphorylated Rab proteins) as a control.
Immunological detection of autophagy and mTORC1-related proteins
By onA general protocol for immunological detection by Western blotting MTOR, MTOR (pS2448), ULK1, ULK1 (pS757), p70S6K, p70S6K (pT389), SQSTM1, CALCOCO2, MAP1LC3B, GABARAP, TFEB, TFE3, PGRN, HSP90, and PCNA.
Mitochondrial isolation protocol
By onThis protocol describes how to isolate crude mitochondrial fractions from HeLa cells.
Immunofluorescence of RAB5 and FLAG-EEA1 puncta after Dynamin-1 and -2 inhibition with Dyngo4a
By onSelective purification of early endosomes can be achieved through affinity capture of the early endosome-associated protein EEA1 (termed Endo-IP) (Park et al. in submission). These purified endosomes can be used for proteomic and lipidomic studies to obtain snapshots of early endosomes. Here we present an immunofluorescence protocol to assess the extent of colocalization between FLAG-EEA1 and RAB5 with and without the Dynamin-1 and -2 (DNM1/2) inhibitor Dyngo4a.
Preparation of unilamellar liposomes
By onA protocol for preparing liposomes that can be used to monitor binding of proteins to vesicles of various membrane curvatures.
Mitophagy induction using Oligomycin/Antimycin A
By onMitophagy induction in HeLa cells using Oligomycin/Antimycin A.
Mitophagy induction using Difereprone
By onProtocol describing the procedure for mitophagy induction in HeLa cells using Difereprone (DFP).
