{"id":11382,"date":"2023-06-07T10:53:22","date_gmt":"2023-06-07T10:53:22","guid":{"rendered":"https:\/\/www.nanion.de\/?post_type=resource_library&#038;p=11382"},"modified":"2023-06-07T10:53:22","modified_gmt":"2023-06-07T10:53:22","slug":"2023-enriched-cell-free-and-cell-based-native-membrane-derived-vesicles-nmv-enabling-rapid-in-vitro-electrophysiological-analysis-of-the-voltage-gated-sodium-channel-1-5","status":"publish","type":"resource_library","link":"https:\/\/nanion.com\/zh-hans\/resource_library\/2023-enriched-cell-free-and-cell-based-native-membrane-derived-vesicles-nmv-enabling-rapid-in-vitro-electrophysiological-analysis-of-the-voltage-gated-sodium-channel-1-5\/","title":{"rendered":"2023 &#8211; Enriched cell-free and cell-based native membrane derived vesicles (nMV) enabling rapid in-vitro electrophysiological analysis of the voltage-gated sodium channel 1.5"},"content":{"rendered":"<p>Here, we demonstrate the utility of native membrane derived vesicles (nMVs) as tools for expeditious electrophysiological analysis of membrane proteins. We used a cell-free (CF) and a cell-based (CB) approach for preparing protein-enriched nMVs. We utilized the Chinese Hamster\u00a0Ovary (CHO) lysate-based cell-free protein synthesis (CFPS) system to enrich ER-derived microsomes in the lysate with the primary human cardiac voltage-gated sodium channel 1.5 (hNa<sub>V<\/sub>1.5; SCN5A) in 3\u00a0h. Subsequently, CB-nMVs were isolated from fractions of nitrogen-cavitated CHO cells overexpressing the hNa<sub>V<\/sub>1.5. In an integrative approach, nMVs were micro-transplanted into\u00a0 <em>Xenopus laevis<\/em>\u00a0oocytes. CB-nMVs expressed native lidocaine-sensitive hNa<sub>V<\/sub>1.5 currents within 24\u00a0h; CF-nMVs did not elicit any response. Both the CB- and CF-nMV preparations evoked single-channel activity on the\u00a0planar lipid bilayer\u00a0while retaining sensitivity to lidocaine application. Our findings suggest a high usability of the quick-synthesis CF-nMVs and maintenance-free CB-nMVs as ready-to-use tools for in-vitro analysis of electrogenic membrane proteins and large, voltage-gated ion channels.<\/p>\n","protected":false},"featured_media":0,"template":"","products_tags":[45,98],"resource_type":[75],"application_areas":[50,53],"therapeutic_areas":[143],"target_categories":[61],"target_families":[332],"targets":[93],"resource_keyword":[],"class_list":["post-11382","resource_library","type-resource_library","status-publish","hentry","products_tags-port-a-patch","products_tags-orbit-16-tc","resource_type-publication","application_areas-ion-channels","application_areas-assay-development-and-validation","therapeutic_areas-assay-development-and-validation","target_categories-voltage-gated-ion-channels","target_families-nav-channels","targets-nav1-5"],"acf":[],"_links":{"self":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_library\/11382","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_library"}],"about":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/types\/resource_library"}],"wp:attachment":[{"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/media?parent=11382"}],"wp:term":[{"taxonomy":"products_tags","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/products_tags?post=11382"},{"taxonomy":"resource_type","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_type?post=11382"},{"taxonomy":"application_areas","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/application_areas?post=11382"},{"taxonomy":"therapeutic_areas","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/therapeutic_areas?post=11382"},{"taxonomy":"target_categories","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/target_categories?post=11382"},{"taxonomy":"target_families","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/target_families?post=11382"},{"taxonomy":"targets","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/targets?post=11382"},{"taxonomy":"resource_keyword","embeddable":true,"href":"https:\/\/nanion.com\/zh-hans\/wp-json\/wp\/v2\/resource_keyword?post=11382"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}