SUIT-009 AmR mt D021

high-resolution terminology - matching measurements at high-resolution

SUIT-009 AmR mt D021

Description

Abbreviation: NS(P)

Reference: A: SUIT-009 short protocol for H₂O₂ (AmR) in mitochondrial preparations (isolated mitochondria and tissue homogenate).

SUIT number: D21_1S;2D;3P;4Rot;5Ama

O2k-Application: AmR

SUIT-category: NS(P)

SUIT protocol pattern: orthogonal 1S;2D;3P;4Rot

SUIT protocols

• [[SUIT-009 AmR mt D21]
• [[SUIT-009 AmR pce D19]
• [[SUIT-009 O2 mt D15]
• [[SUIT-009 O2 pce D16]

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 UltraRed® 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 °C.

For O2-Application, choose SUIT-9 O2 mt D15; for O2 and H₂O₂ measurements (AmR), choose: D21_1S;2D;3P;4Rot;5Ama.

MitoPedia concepts: MiP concept, SUIT protocol, SUIT A, Recommended, Find 

MitoPedia methods: Fluorometry