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SUIT-009 AmR ce-pce D019

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Revision as of 10:48, 17 January 2019 by Iglesias-Gonzalez Javier (talk | contribs) (Iglesias-Gonzalez Javier moved page SUIT-009 AmR pce D19 to SUIT-009 AmR pce D019: New Nomenclature)


high-resolution terminology - matching measurements at high-resolution


SUIT-009 AmR ce-pce D019

Description

SUIT9

Abbreviation: NS(P)

Reference: A: SUIT-009 short protocol for H2O2 (AmR) in pce.

SUIT number: D019_1S;2D;3P;4Rot;5Ama

O2k-Application: AmR

SUIT-category: NS(P)
SUIT protocol pattern: orthogonal 1S;2D;3P;4Rot
SUIT-9 AmR pce D19 Overview.png

1S;2D;3P;4Rot;5Ama-

SUIT 9 is a short protocol for simultaneous determination of O2 flux and the rate of H2O2 production (H2O2 flux) on isolated mitochondria,tissue homogenate (except of liver) and permeabilized cells. Succinate (S) supports the reverse electron transfer (RET) initiated H2O2 production in the LEAK state. Addition of ADP decreases H2O2 flux due to depolarization of the mitochondrial membrane. Pyruvate (P) supports NADH-pathway (N), and usually in the presence of ADP it does not increase further the H2O2 flux. Antimycin A (Ama) inhibits Complex III (CIII) and results in the elevation of the H2O2 flux. The sensitivity of the Amplex UltraRedยฎ assay (for determining H2O2 production) is changing over the experimental time and upon addition of chemicals. To correct H2O2 flux for the sensitivity changes several H2O2 calibration steps are done during the experiment. The measurement is carried out in MiR05-Kit at 37 ยฐC.


MitoPedia concepts: SUIT protocol, SUIT A, Find 


MitoPedia methods: Fluorometry