Vesicle isolation - somebody know a protocol for that? (May/22/2007 )

Hi everybody,

I'll like to know if somebody knows about a protocol for isolation of vesicles, Im intrested in endositic vesicles in general for I want to look for a membrain atacched receptor endocited after inactivation.

Any help or reference could help

Thanks!!

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Check this paper. 'Molecular Anatomy of a Trafficking Organelle'. I think they have described synaptic vesicle isolation.

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vesicle isolation is hard work; each tissue (exocrine, endocrine, neuroendocrine or neuronal and others) have own protocols; grab articles of the leader in the field who set the current "state of the art" method, and adapt it; there are even for same tissues various protocols around but not all are generally accepted;

common techniques are homogeneization of tissue or cells (not lysis), sucrose gradient density centrifugation and immunoisolation; you should combine various techniques

think of monitoring your enrichment by measuring reference enzymes or proteins, especially to describe the grade of contamination with other membranes (organelles or PM)

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Thanks a lot!!!

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