Difference between revisions of "SUIT-009 O2 ce-pce D016"

Revision as of 13:20, 21 January 2019

high-resolution terminology - matching measurements at high-resolution

SUIT-009 O2 ce-pce D016

Description

Abbreviation: NS(P)

Reference: A: SUIT-009 short protocol for mt-respiration in pce.

SUIT number: D016_ce1;1Dig;1S;2D;3P;4Rot;5Ama

O2k-Application: O2

SUIT-category: NS(P)

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

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

SUIT-009 is a short protocol for determination of O₂ flux on permeabilized cells. Succinate (S) supports the reverse electron transfer (RET) initiated H₂O₂ production in the LEAK state. Addition of ADP increases S-OXPHOS capacity. Pyruvate (P) supports NADH-pathway (N). Antimycin A (Ama) inhibits Complex III (CIII) and results in the inhibition of O₂ flux. For H2O2-Application, choose SUIT-009 AmR pce D019; for O2 and H₂O₂ measurements (AmR), choose: SUIT-009 AmR pce D019.

MitoPedia concepts: SUIT protocol, SUIT A, Find

MitoPedia methods: Fluorometry

@@ Line 2: / Line 2: @@
 |abbr=NS(P)
 |description=[[File:1S;2D;3P;4Rot-.png|400px|SUIT-009]]
-|info='''A''': '''[[SUIT-009]]''' short protocol for H<sub>2</sub>O<sub>2</sub> (AmR) in [[permeabilized cells |pce]].
+|info='''A''': '''[[SUIT-009]]''' short protocol for mt-respiration  in [[permeabilized cells |pce]].
 |application=O2
 |SUIT number=D016_ce1;1Dig;1S;2D;3P;4Rot;5Ama
@@ Line 12: / Line 12: @@
 == 1S;2D;3P;4Rot;5Ama-==
-SUIT-009 is a short protocol for simultaneous determination of O<sub>2</sub> flux and the rate of [[MiPNet20.14 AmplexRed H2O2-production|H<sub>2</sub>O<sub>2</sub> production]] (H<sub>2</sub>O<sub>2</sub> flux) on isolated mitochondria,tissue homogenate (except of liver) and permeabilized cells. Succinate (S) supports the reverse electron transfer (RET) initiated H<sub>2</sub>O<sub>2</sub> production in the LEAK state. Addition of ADP decreases H<sub>2</sub>O<sub>2</sub> flux due to depolarization of the mitochondrial membrane. Pyruvate (P) supports [[NADH Electron transfer-pathway state|NADH-pathway (N)]], and usually in the presence of ADP it does not increase further the H<sub>2</sub>O<sub>2</sub> flux. Antimycin A (Ama) inhibits Complex III (CIII) and results in the elevation of the H<sub>2</sub>O<sub>2</sub> flux. The sensitivity of the Amplex UltraRed® assay (for determining H<sub>2</sub>O<sub>2</sub> production) is changing over the experimental time and upon addition of chemicals. To correct H<sub>2</sub>O<sub>2</sub> flux for the sensitivity changes several [[MiPNet20.14_AmplexRed_H2O2-production#Calibration_with_H2O2|H<sub>2</sub>O<sub>2</sub> calibration steps]] are done during the experiment. The measurement is carried out in MiR05-Kit at 37 °C.
+SUIT-009 is a short protocol for determination of O<sub>2</sub> flux  on permeabilized cells. Succinate (S) supports the reverse electron transfer (RET) initiated H<sub>2</sub>O<sub>2</sub> production in the LEAK state. Addition of ADP increases S-OXPHOS capacity. Pyruvate (P) supports [[NADH Electron transfer-pathway state|NADH-pathway (N)]]. Antimycin A (Ama) inhibits Complex III (CIII) and results in the inhibition of O<sub>2</sub> flux.
+For H2O2-Application, choose [[SUIT-009 AmR pce D019]]; for O2 and H<sub>2</sub>O<sub>2</sub> measurements (AmR), choose: [[SUIT-009 AmR pce D019]].
 {{MitoPedia concepts