Cell Biology

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    Protocols in Current Issue
    Cell-free Reconstitution of the Packaging of Cargo Proteins into Vesicles at the trans Golgi Network
    Authors:  Xiao Tang, Feng Yang and Yusong Guo, date: 03/05/2020, view: 1036, Q&A: 0
    [Abstract] Protein sorting at the trans Golgi network (TGN) plays important roles in targeting newly synthesized proteins to their specific destinations. The aim of this proposal is to reconstitute the packaging of non-Golgi resident cargo proteins into vesicles at the TGN, utilizing rat liver cytosol, semi-intact mammalian cells and nucleotides. ...
    Cell-free Fluorescent Intra-Golgi Retrograde Vesicle Trafficking Assay
    Authors:  Nathanael P. Cottam and Daniel Ungar, date: 11/20/2017, view: 4565, Q&A: 0
    [Abstract] Intra-Golgi retrograde vesicle transport is used to traffic and sort resident Golgi enzymes to their appropriate cisternal locations. An assay was established to investigate the molecular details of vesicle targeting in a cell-free system. Stable cell lines were generated in which the trans-Golgi enzyme galactosyltransferase (GalT) was ...
    Preparation of Golgi Membranes from Rat Liver
    Authors:  Carmen Valente, Gabriele Turacchio, Stefania Spanò, Alberto Luini and Daniela Corda, date: 11/20/2014, view: 6416, Q&A: 0
    [Abstract] This protocol details the isolation of enriched Golgi membranes from rat liver, using discontinuous density gradient centrifugation. This high-yield extraction method is useful for several applications, including immunoprecipitation of solubilised Golgi membrane proteins (preparation included) and electron microscopy. Protocol adapted from ...
    Enrichment of Golgi membranes from HeLa cells by sucrose gradient ultracentrifugation
    Authors:  Josse van Galen and Julia von Blume, date: 09/20/2013, view: 11369, Q&A: 1
    [Abstract] This is a protocol to extract intact Golgi Membranes from HeLa cells using sucrose gradient centrifugation. This extraction is very useful for several applications including pull-down of Golgi membrane proteins, electron microscopy and reconstitution of protein transport into an isolated system. Protocol adapted from Balch et al. (1984).
    Subcellular Fractionation Using Accudenz Gradient to Separate ER/Golgi in Yeast
    Author:  Zongtian Tong, date: 01/20/2012, view: 13553, Q&A: 0
    [Abstract] This protocol describes how to separate the endoplasmic reticulum (ER) and Golgi apparatus in yeast cells using a subcellular fractionation approach with an Accudenz gradient.



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