Description
Abbreviation: RET (respiratory control) of SUIT-026 AmR ce-pce D087
Reference: B - SUIT-026 - Protocol for the respiratory control of SUIT-026 AmR ce-pce D087
SUIT number: D088_ce1;1Dig;1S;2Rot;3D;3c;4Ama
O2k-Application: O2
in preparation
Communicated by Cardoso LHD (last update 2022-09-05)
Representative traces
SUIT-026 O2 ce-pce D088 has been designed to serve as a respiratory control of the SUIT-026 AmR ce-pce D087 (protocol to study the RET-initiated ROS production in permeabilized cells (adding living cells into the chamber and permeabilizing the plasma membrane with digitonin titration.
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 ce-pce D088.
- + 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
- SUIT-026 AmR ce-pce D087: Protocol to evaluate the RET production of ROS.
- SUIT-006 O2 mt D022: CCP mtprep for S-pathway.
Chemicals and syringes
Template:Chemicals SUIT-026 O2 ce-pce D088
- Suggested stock concentrations are shown in the specific DL-Protocol.
References
MitoPedia concepts: SUIT protocol, SUIT B, Find
MitoPedia methods:
Fluorometry