Abbreviation: RET (respiratory control) of SUIT-026 AmR mt D064
SUIT number: D063_1S;2Rot;3D;3c;4Ama
SUIT-026 O2 mt D063 has been designed to serve as a respiratory control of the SUIT-026 AmR mt D064 (protocol to study the RET-initiated ROS production in mitochondrial preparations (isolated mitochondria, tissue homogenate and permeabilized cells which are already permeabilized when they are added to the chamber).
Communicated by Komlodi T, Iglesias-Gonzalez J and Gnaiger E (last update 2019-09-24)
Steps and respiratory states
|Step||State||Pathway||Q-junction||Comment - Events (E) and Marks (M)|
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Pathway control
Strengths and limitations
- This protocol serves as a respiratory control (since fluorescence dyes may inhibit components of the ET system) and quality control for the sample for SUIT-026 AmR mt D064.
- + Short protocol.
- + The addition of cytochrome c to assess the mt outer membrane integrity provides a quality control of the sample, which cannot be performed in the protocol with Amplex UltraRed.
- - This protocol does not provide information about all the coupling control states (LEAK, OXPHOS and ET). However, it is possible to create a DLPU by additing an Event&Mark for the uncoupler titration (4U) after ADP (3D) to obtain ET-state.
Compare SUIT protocols
Chemicals and syringes
|Step||Chemical(s) and link(s)||Comments|
|3c||Cytochrome c (c)|
|4Ama||Antimycin A (Ama)|
- Suggested stock concentrations are shown in the specific DL-Protocol.
MitoPedia methods: Fluorometry