Fractionation of subcellular membrane vesicles of epithelial and non-epithelial cells by OptiPrep™ density gradient ultracentrifugation

Xuhang Li; Mark Donowitz

doi:10.1007/978-1-4939-0944-5_6

Fractionation of subcellular membrane vesicles of epithelial and non-epithelial cells by OptiPrep™ density gradient ultracentrifugation

Xuhang Li, Mark Donowitz

School of Medicine

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

Density gradient ultracentrifugation (DGUC) is widely used for physical isolation (enrichment rather than purifi cation) of subcellular membrane vesicles. It has been a valuable tool to study specifi c subcellular localization and dynamic traffi cking of proteins. While sucrose has been the main component of density gradients, several years ago, synthetic OptiPrep™ (iodixanol) began being used for separation of organelles due to its iso-osmotic property. Here, we describe a detailed protocol for density gradient fractionation of various mammalian subcellular vesicles, including endoplasmic reticulum (ER), Golgi apparatus, endosomes, and lipid rafts, as well as apical and basolateral membranes of polarized epithelial cells.

Original language	English (US)
Pages (from-to)	85-99
Number of pages	15
Journal	Methods in Molecular Biology
Volume	1174
DOIs	https://doi.org/10.1007/978-1-4939-0944-5_6
State	Published - 2014

Keywords

Density gradient ultracentrifugation
Epithelia
Lipid rafts
OptiPrep
Organelles
Subcellular membrane vesicles

ASJC Scopus subject areas

Molecular Biology
Genetics

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10.1007/978-1-4939-0944-5_6

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abstract = "Density gradient ultracentrifugation (DGUC) is widely used for physical isolation (enrichment rather than purifi cation) of subcellular membrane vesicles. It has been a valuable tool to study specifi c subcellular localization and dynamic traffi cking of proteins. While sucrose has been the main component of density gradients, several years ago, synthetic OptiPrep{\texttrademark} (iodixanol) began being used for separation of organelles due to its iso-osmotic property. Here, we describe a detailed protocol for density gradient fractionation of various mammalian subcellular vesicles, including endoplasmic reticulum (ER), Golgi apparatus, endosomes, and lipid rafts, as well as apical and basolateral membranes of polarized epithelial cells.",

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T1 - Fractionation of subcellular membrane vesicles of epithelial and non-epithelial cells by OptiPrep™ density gradient ultracentrifugation

AU - Li, Xuhang

AU - Donowitz, Mark

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PY - 2014

Y1 - 2014

N2 - Density gradient ultracentrifugation (DGUC) is widely used for physical isolation (enrichment rather than purifi cation) of subcellular membrane vesicles. It has been a valuable tool to study specifi c subcellular localization and dynamic traffi cking of proteins. While sucrose has been the main component of density gradients, several years ago, synthetic OptiPrep™ (iodixanol) began being used for separation of organelles due to its iso-osmotic property. Here, we describe a detailed protocol for density gradient fractionation of various mammalian subcellular vesicles, including endoplasmic reticulum (ER), Golgi apparatus, endosomes, and lipid rafts, as well as apical and basolateral membranes of polarized epithelial cells.

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KW - Epithelia

KW - Lipid rafts

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KW - Organelles

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Fractionation of subcellular membrane vesicles of epithelial and non-epithelial cells by OptiPrep™ density gradient ultracentrifugation

Abstract

Keywords

ASJC Scopus subject areas

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