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- W2895877943 abstract "During in vitro homotypic yeast vacuole fusion Ca2+ is transported into and out of the organelle lumen. In vitro, Ca2+ is taken up from the medium by vacuoles upon the addition of ATP. During the docking stage of vacuole fusion Ca2+ is effluxed from the lumen upon the formation of trans-SNARE complexes between vesicles. Here we describe a real-time fluorescence-based assay to monitor the transport of this cation using purified organelles. Extraluminal Ca2+ is detected when the cation binds the low-affinity fluorescent dye Fluo-4 dextran. This allows for the use of a 96-well microtiter plate to be read in a fluorescence plate reader. Thus, in addition to a curve of calibrated Ca2+ standards, up to 91 experimental conditions can be monitored in a single microplate using this method." @default.
- W2895877943 created "2018-10-26" @default.
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- W2895877943 date "2018-10-14" @default.
- W2895877943 modified "2023-09-27" @default.
- W2895877943 title "Real-Time Fluorescence Detection of Calcium Efflux During Vacuolar Membrane Fusion" @default.
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- W2895877943 doi "https://doi.org/10.1007/978-1-4939-8760-3_21" @default.
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