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One‐step separation of endocytic organelles, Golgi/ trans ‐Golgi network and plasma membrane by density gradient electrophoresis
Author(s) -
Lindner Robert
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200102)22:3<386::aid-elps386>3.0.co;2-t
Subject(s) - golgi apparatus , endosome , organelle , cell fractionation , endocytic cycle , microbiology and biotechnology , chemistry , membrane , free flow electrophoresis , biology , endoplasmic reticulum , biochemistry , endocytosis , cell , polyacrylamide gel electrophoresis , gel electrophoresis of proteins , enzyme
Many different methods for the fractionation of subcellular organelles have been reported. However, no protocol for rapid separation of plasma membrane, Golgi/ trans ‐Golgi network (TGN) and endosomes is available to date. Such a method is a prerequisite for a quantitative biochemical analysis of vesicular transport from the Golgi/TGN compartment to plasma membrane and endosomes. Here a density gradient electrophoresis protocol is described that allows the fractionation of these organelles in one step. This protocol requires only low‐cost instrumentation available in most biochemical laboratories.