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SUIT-026 O2 ce-pce D088

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SUIT-026 O2 ce-pce D088

Description

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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

Template:SUIT text D088

Communicated by  Cardoso LHD (last update 2022-09-05)


Representative traces

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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


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