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Mitochondrial Bioenergetics
Carlos, M Palmeira [szerk.]
;
António, J Moreno [szerk.]
Angol nyelvű Szakkönyv (Könyv) Tudományos
Megjelent: Springer Science+Business Media, New York, Amerikai Egyesült Államok, 19 p.
2018
Sorozatok:
Methods in Molecular Biology 1064-3745 1940-6029, 1782
Azonosítók
MTMT: 3407055
ISBN:
9781493978304
High-Resolution FluoRespirometry is a well-established and versatile approach to study mitochondrial oxygen uptake amperometrically in combination with measurement of fluorescence signals. One of the most frequently applied fluorescent dyes is Amplex UltraRed for monitoring rates of hydrogen peroxide production. Selection of an appropriate mitochondrial respiration medium is of crucial importance, the primary role of which is to support and preserve optimum mitochondrial function. For harmonization of results in a common database, we compared respiration and H2O2 production of permeabilized HEK 293T cells measured in MiR05 (sucrose and K-lactobionate), Buffer Z (K-MES and KCl), MiR07 (combination of MiR05 and Buffer Z), and MiRK03 (KCl). Respiration in a simple substrate-uncoupler-inhibitor titration protocol was identical in MiR05, Buffer Z, and MiR07, whereas oxygen fluxes detected with MiRK03 were consistently lower in all coupling and electron transfer-pathway states. H2O2 production rates were comparable in all four media, while assay sensitivity was comparatively low with MiR05 and MiR07 and higher but declining over time in the other two media. Stability of assay sensitivity over experimental time was highest in MiR05 but slightly less in MiR07. Taken together, MiR05 and Buffer Z yield comparable results on respiration and H2O2 production. Despite the lower sensitivity, MiR05 was selected as the medium of choice for FluoRespirometry due to the highest stability of the sensitivity or calibration constant observed in experiments over periods of up to 2 h. © 2018, Springer Science+Business Media, LLC, part of Springer Nature.
Fejezetek
Komlódi T et al. Comparison of mitochondrial incubation media for measurement of respiration and hydrogen peroxide production. (2018) Megjelent: Mitochondrial Bioenergetics pp. 137-155
Kurz F.T. et al. Assessing spatiotemporal and functional organization of mitochondrial networks. (2018) Megjelent: Mitochondrial Bioenergetics pp. 383-402
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2025-04-26 07:43
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