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Rekonstituering af et transmembranprotein, Voltage ionkanal, KvAP, i Giant unilamelvesikler til mikroskopi og Patch clamp-forsøg

DOI :

10.3791/52281-v

January 22nd, 2015

January 22nd, 2015

18,682 Views

1Institut Curie, Centre de Recherche, CNRS, UMR 168, PhysicoChimie Curie, Université Pierre et Marie Curie, 2Kavli Institute for Brain and Mind, University of California, San Diego, 3Molecular Physiology and Biophysics Section, National Institute for Neurological Disorders and Stroke, National Institute of Health

Genoprettelse af det transmembrane protein, KvAP, i gigantiske unilamelblærer (GUVs), er dokumenteret i to dehydrering-rehydrering metoder - electroformation og gel-assisteret hævelse. I begge fremgangsmåder er små unilamellare vesikler indeholdende proteinet fusioneret sammen for at danne GUVs, som derefter kan studeres ved fluorescensmikroskopi og patch-clamp elektrofysiologi.

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