What should be the best protocol for plasma membrane preparation from cultured mammalian cell lines ?
My goal is to obtain large (micron) sheets of plasma membranes in their native state, for electron microscopy and cryomicroscopy.
Thanks for sharing your ideas.
Pierre,
Je prépare des feuillets de membrane plasmique de cellules chromaffines pour faire des doubles marquages. La technique est décrite dans le papier Umbrecht-Jenck et al 2010 dans Traffic. Si tu veux plus d'infos tu peux m'appeler.
Cordialement
Sylvette
Pierre,
Try the following paper:
http://www.ncbi.nlm.nih.gov/pubmed/21701974
Hope this helps!
Cheers,
Matt
Hi Pierre!
I think, the best way to observe large membrane sheets is Freeze-fracture! After Freeze-fracture you can look on the membrane from above and additionally you can label proteins or lipids you are interested in. But thats definitely not native. Depending how you fix the cells the preparation could be very close to the native stages.
Best,
Eric
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