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  A convenient protocol for generating giant unilamellar vesicles containing SNARE proteins using electroformation.

Witkowska, A., Jablonski, L., & Jahn, R. (2018). A convenient protocol for generating giant unilamellar vesicles containing SNARE proteins using electroformation. Scientific Reports, 8: 9422. doi:10.1038/s41598-018-27456-4.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0001-9C55-0 Version Permalink: http://hdl.handle.net/21.11116/0000-0003-CA64-9
Genre: Journal Article

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Witkowska, A.1, Author              
Jablonski, L., Author
Jahn, R.1, Author              
Affiliations:
1Department of Neurobiology, MPI for Biophysical Chemistry, Max Planck Society, ou_578595              

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 Abstract: Reconstitution of membrane proteins in artificial membranes is an essential prerequisite for functional studies that depend on the context of an intact membrane. While straight-forward protocols for reconstituting proteins in small unilamellar vesicles were developed many years ago, it is much more difficult to prepare large membranes containing membrane proteins at biologically relevant concentrations. Giant unilamellar vesicles (GUVs) represent a model system that is characterised by low curvature, controllable tension, and large surface that can be easily visualised with microscopy, but protein insertion is notoriously difficult. Here we describe a convenient method for efficient generation of GUVs containing functionally active SNARE proteins that govern exocytosis of synaptic vesicles. Preparation of proteo-GUVs requires a simple, in-house-built device, standard and inexpensive electronic equipment, and employs a straight-forward protocol that largely avoids damage of the proteins. The procedure allows upscaling and multiplexing, thus providing a platform for establishing and optimizing preparation of GUVs containing membrane proteins for a diverse array of applications.

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Language(s): eng - English
 Dates: 2018-06-21
 Publication Status: Published online
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 Rev. Method: Peer
 Identifiers: DOI: 10.1038/s41598-018-27456-4
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Title: Scientific Reports
Source Genre: Journal
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Pages: 8 Volume / Issue: 8 Sequence Number: 9422 Start / End Page: - Identifier: -