體外熒光法檢測核內體早期動力學2
Full size image (70 KB)In vitro incubation of a reaction mix that contains labeled endosomes, cytosol and an ATP-regenerating system at physiological temperature leads to both docking and fusion, or sorting and budding of the endosomes (Steps 31–39). Depending on the combination of labeled endosomes used, either docking/fusion or sorting/budding is preferentially visualized. The purpose of the assay is to t......閱讀全文
體外熒光法檢測核內體早期動力學-2
Full size image (70?KB)In vitro?incubation of a reaction mix that contains labeled endosomes, cytosol and an ATP-regenerating system at physiological
體外熒光法檢測核內體早期動力學
A fluorescence-based?in vitro?assay for investigating early endosome dynamicsSina V Barysch1,2, Reinhard Jahn1?& Silvio O Rizzoli2ABSTRACTEarly endoso
體外熒光法檢測核內體早期動力學-3
REAGENT SETUP Critical step?All solutions and buffers must be free of detergent and therefore should be prepared in plasticware, or in glassware th
體外熒光法檢測核內體早期動力學-6
Critical step?Be careful to thaw PNS and cytosol slowly on ice to avoid unwanted protein degradation or breaking of organelles. This might require
體外熒光法檢測核內體早期動力學-5
Leave tubes on ice and repeat Steps 13–15 for the next 6–12 plates of cells.When all cells are collected, centrifuge all tubes at 250g?for 5 min at 4
體外熒光法檢測核內體早期動力學-4
Top?of pageProcedureOverviewSteps 1 - 8 Preparation of rat brain cytosolSteps 9 - 10 Culture of PC12 cellsSteps 11 - 30 Preparation of postnuclear sup
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