Extracting only plasma membrane - (May/20/2008 )

My PI wants me to grow up some human cancer cells in culture and then collect their plasma membranes for someone else to run tests on. All the protocols I've found so far either focus on getting rid of the plasma membrane or require actual tissue. Any ideas on how to collect just plasma membrane?

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Check these links:
http://search.vadlo.com/b/q?sn=158621799&a...ation&rel=0
..

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Qiagen has a kit that extracts proteins from 4 cell parts in 4 steps: cytoplasm, cytoskeleton, plasma membrane and nucleus. it is famous. find from the catalog.

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As best I can tell, my PI doesn't just want the proteins, but the membranes as whole as possible.

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...then your PI is crazy...

but actually the link that someone gave above is good, I checked the links, the protocols are quite alright.

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I doubt that one can isolate plasma membrane w/o contamination form other organelle membranes with simple protocols;

what can be done is to enrich by sucrose density centrifugation and subsequent immunprecipitation against a surface protein that is unique to the plasmam membrane

anyway, if using kits or self-made protocols, you have to check recovery and enrichment factor by measuring reference proteins (enzymes) for each relevant membrane-bearing organelle and the plasma membrane

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