Template:SUIT text D034: Difference between revisions
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SUIT-006 Fluo mt D034 is a protocol to investigate the O<sub>2</sub> flux and [[mitochondrial membrane potential]] with fluorescent dyes. In this protocol, the [[NADH Electron transfer-pathway state]] can be analysed [[Mitochondrial preparations| mitochondrial preparations]] such as isolated mitochondria, tissue homogenates and permeabilized cells (already permeabilized when they are added to the chamber) in a wide variety of species and tissues. | SUIT-006 Fluo mt D034 is a protocol to investigate the O<sub>2</sub> flux and [[mitochondrial membrane potential]] with fluorescent dyes. In this protocol, the [[NADH Electron transfer-pathway state]] can be analysed in [[Mitochondrial preparations| mitochondrial preparations]] such as isolated mitochondria, tissue homogenates and permeabilized cells (already permeabilized when they are added to the chamber) in a wide variety of species and tissues. | ||
Addition of PM ([[pyruvate]] & [[malate]]) to the mitochondria leads to the hyperpolarisation of the mt-membrane, while [[ADP]] (D) decreases the mt-membrane potential. Addition of [[oligomycin]] (Omy) results in hyperpolarisation since the inhibition of the [[ATP synthase]] leads to an accumulation of protons in the intermembrane space. [[Uncoupler]] depolarises the mt-membrane in a concentration-dependent manner and [[antimycin A]] blocks the respiration and dissipates the mt-membrane potential. | Addition of PM ([[pyruvate]] & [[malate]]) to the mitochondria leads to the hyperpolarisation of the mt-membrane, while [[ADP]] (D) decreases the mt-membrane potential. Addition of [[oligomycin]] (Omy) results in hyperpolarisation since the inhibition of the [[ATP synthase]] leads to an accumulation of protons in the intermembrane space. [[Uncoupler]] depolarises the mt-membrane in a concentration-dependent manner and [[antimycin A]] blocks the respiration and dissipates the mt-membrane potential. | ||
Since high concentrations of Omy can decrease the ET-capacity induced by addition of uncoupler, the optimal concentration of Omy has to be determined. | Since high concentrations of Omy can decrease the ET-capacity induced by addition of uncoupler, the optimal concentration of Omy has to be determined. | ||
Revision as of 15:55, 19 July 2019
SUIT-006 Fluo mt D034 is a protocol to investigate the O2 flux and mitochondrial membrane potential with fluorescent dyes. In this protocol, the NADH Electron transfer-pathway state can be analysed in mitochondrial preparations such as isolated mitochondria, tissue homogenates and permeabilized cells (already permeabilized when they are added to the chamber) in a wide variety of species and tissues. Addition of PM (pyruvate & malate) to the mitochondria leads to the hyperpolarisation of the mt-membrane, while ADP (D) decreases the mt-membrane potential. Addition of oligomycin (Omy) results in hyperpolarisation since the inhibition of the ATP synthase leads to an accumulation of protons in the intermembrane space. Uncoupler depolarises the mt-membrane in a concentration-dependent manner and antimycin A blocks the respiration and dissipates the mt-membrane potential. Since high concentrations of Omy can decrease the ET-capacity induced by addition of uncoupler, the optimal concentration of Omy has to be determined.
