SUIT-009 O2 mt D015

high-resolution terminology - matching measurements at high-resolution

SUIT-009 O2 mt D015

Description

SUIT9

Abbreviation: NS(P)

Reference: A: SUIT 9 is a short SUIT protocol for H₂O₂ (AmR) in mitochondrial preparations.

SUIT number: 9

O2k-Application: AmR

MitoPedia concepts: SUIT protocol, SUIT B 

SUIT-category: NS(P)

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

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.

SUIT O2 DLP for SUIT 9 (DatLab 7.1)

DL-Protocols (DLP) for substrate-uncoupler-inhibitor titrations (see MitoPedia: SUIT) guide through a sequence of coupling and pathway control states. A specific SUIT protocol can be assigned to O2k-Chamber A or B or both. DL-Protocols for different SUIT protocols can be found in the library of protocols MitoPedia: SUIT. Instrumental DL-protocols are displayed in the DL-Protocol library.

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DL-Protocol	Description	Download - Protocol .DLP	Download - Analysis .XLSX
AmR_SUIT 9_mt	SUIT protocol for H2O2 (AmR) in mitochondrial preparations (imt, thom)
AmR_SUIT 9_pce	SUIT protocol for H2O2 (AmR) in permeabilized cells (pce)