SUIT-009 O2 mt D015: Difference between revisions

Revision as of 15:29, 13 June 2018

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 A

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 7.3)

Updating this page (2018-06-13) for DatLab 7.3 DLP files

@@ Line 20: / Line 20: @@
 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.
-== SUIT O2 DLP for SUIT 9 (DatLab 7.1) ==
+== SUIT O2 DLP for SUIT 9 (DatLab 7.3) ==
-:::: [[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-Protocols#DL-Protocol_library| DL-Protocol library]].
+''Updating this page (2018-06-13) for DatLab 7.3 DLP files''
-{{Template:Download instructions}}
-{| class="wikitable" border="1"
-|-
-! DL-Protocol
-! Description
-! Download - Protocol .DLP
-! Download - Analysis .XLSX
-|-
-| AmR_SUIT 9_mt
-| SUIT protocol for H<sub>2</sub>O<sub>2</sub> (AmR) in mitochondrial preparations (imt, thom)
-| style="text-align:center"| [[File:Download1.png|20px|link=http://www.bioblast.at/images/c/ca/AmR_imt_thom_1S%3B2D%3B3P%3B4Ama.DLP]]
-|
-|-
-| AmR_SUIT 9_pce
-| SUIT protocol for H<sub>2</sub>O<sub>2</sub> (AmR) in permeabilized cells (pce)
-| style="text-align:center"| [[File:Download1.png|20px|link=http://www.bioblast.at/images/5/59/AmR_pce_1S%3B2D%3B3P%3B4Ama.DLP]]
-|
-|}