A Microplate-Based Bioluminescence Assay of Mitochondrial Calcium Uptake
| Autor: | Javier García-Sancho, Maria Teresa Alonso, Paloma Navas-Navarro |
|---|---|
| Přispěvatelé: | Instituto de Biología y Genética Molecular (IBGM), Universitad de Valladolid, IBGM, University of Valladolid |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
MESH: Mitochondria Aequorin Mitochondrion 03 medical and health sciences Bioluminescence Mitochondrial calcium uptake MESH: Statistics as Topic MESH: Humans Calcium uniporter MESH: Aequorin biology Chemistry [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biology MESH: Luminescent Measurements 030104 developmental biology Apoptosis MESH: Calcium MESH: HeLa Cells Screening MESH: Calcium Channels biology.protein Biophysics Cell activation Homeostasis Plate reader Plate-reader |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493968220 Methods in Molecular Biology Methods in Molecular Biology, 1567, pp.245-253, 2017, ⟨10.1007/978-1-4939-6824-4_15⟩ |
| DOI: | 10.1007/978-1-4939-6824-4_15 |
| Popis: | International audience; Mitochondrial Ca2+ homeostasis is crucial for regulating vital functions such as respiration or apoptosis. Targeted aequorins are excellent probes to measure subcellular Ca2+. Ca2+ concentration in mitochondria ([Ca2+]M) is low at rest (about 10-7 M) and can increase to the micromolar or even approach the millimolar range, upon cell activation. Here we describe a new quantitative luminescent protocol to directly measure mitochondrial Ca2+ uptake, optimized for high throughput. The sensitivity of the method allows detection of changes in either the capacity or the affinity of mitochondrial Ca2+ transport. |
| Databáze: | OpenAIRE |
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