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Isolating HeLa cell fractions enriched for clathrin-coated vesicles
Georg H H Borner1, Andrew B Fielding2
1Department of Proteomics and Signal Transduction, Max Plank Institute of Biochemistry, 82152 Martinsried, Germany;
Cold Spring Harbor Protocols
|November 5, 2014
Summary
This study details a protocol for isolating clathrin-coated vesicles (CCVs) from HeLa cells. This method allows researchers to analyze how drugs and gene silencing affect CCV composition.
Area of Science:
- Cell biology
- Molecular and cellular biology
Background:
- HeLa cell lines are widely used for experimental manipulation.
- Clathrin-coated vesicles (CCVs) are crucial for intracellular trafficking.
- Understanding CCV composition is vital for studying cellular processes.
Purpose of the Study:
- To describe a protocol for isolating CCV-enriched fractions from HeLa cells.
- To provide a method for analyzing the impact of experimental manipulations on CCV composition.
Main Methods:
- Utilizes HeLa cell lines for experimental manipulation.
- Employs drug treatments or RNA interference for gene silencing.
- Isolates fractions enriched for clathrin-coated vesicles (CCVs).
Main Results:
- Successfully prepared a HeLa cell fraction enriched for a mixed population of CCVs.
- The isolated CCV fraction is suitable for downstream analyses.
Conclusions:
- The described protocol enables the study of CCV composition changes.
- This method facilitates research into the effects of cellular manipulations on CCVs.
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