SUIT-009 O2 ce-pce D016: Difference between revisions

Revision as of 10:40, 15 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 H₂O₂ (AmR) in pce.

SUIT number: D16_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 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.

MitoPedia concepts: SUIT protocol, SUIT A

MitoPedia methods: Fluorometry

@@ Line 1: / Line 1: @@
 {{MitoPedia
 |abbr=NS(P)
+|description=[[File:1S;2D;3P;4Rot-.png|400px|SUIT9]]
+|info='''A''': '''[[SUIT-009]]''' short protocol for H<sub>2</sub>O<sub>2</sub> (AmR) in [[permeabilized cells |pce]].
+|application=O2
 |SUIT number=D16_ce1;1Dig;1S;2D;3P;4Rot;5Ama
-|application=O2
-|description=[[File:1S;2D;3P;4Rot-.png|400px|SUIT9]]
-|info='''A''': '''SUIT 9''' is a short SUIT protocol for H<sub>2</sub>O<sub>2</sub> (AmR) in permeabilized cells.
-}}
-{{MitoPedia concepts
-|mitopedia concept=SUIT protocol, SUIT A
 }}
 ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS(P)
+::: '''[[SUIT protocol pattern]]:''' orthogonal 1S;2D;3P;4Rot
 __TOC__
@@ Line 19: / Line 13: @@
 == 1S;2D;3P;4Rot;5Ama-==
 SUIT 9 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.
+{{MitoPedia concepts
+|mitopedia concept=SUIT protocol, SUIT A
+}}
+{{MitoPedia methods
+|mitopedia method=Fluorometry
+}}