SUIT-003 O2 ce D061: Difference between revisions
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{{MitoPedia | {{MitoPedia | ||
|abbr=ce | |abbr=CCP-ce Snv,Mnanv - control | ||
|description=[[File: | |description=[[File:ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama.png|400px]] | ||
|info='''B''' | |info='''B''' Carrier control for the [[coupling-control protocol]] for [[living cells]] with cell permeable compounds [[MitoKit-CII/Succinate-nv]] and [[MitoKit-CII/Malonate-nv]] - [[SUIT-003 O2 ce D060]] | ||
|application=O2 | |application=O2 | ||
|SUIT number=D061_ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama | |||
}} | }} | ||
::: '''[[SUIT protocol pattern]]:''' | ::: '''[[MitoPedia: SUIT]]''' - Coupling-control protocol (CCP) with the carrier of the plasma membrane-permeable succinate ([[MitoKit-CII/Succinate-nv]]) and plasma membrane-permeable malonate ([[MitoKit-CII/Malonate-nv]]) in living cells (ce): DMSO. | ||
::: '''[[SUIT protocol pattern]]:''' ce1;(ce2Omy);ce3U;ce4DMSO;ce5DMSO;ce6Rot;ce7Ama | |||
{{Template:SUIT text D061}} | |||
SUIT-003 O2 ce D061 protocol is a control for the [[SUIT-003 O2 ce D060]], with the addition of the [[Carrier control titrations|carrier]] DMSO instead of the plasma membrane-permeable succinate ([[[MitoKit-CII/Succinate-nv]]) and plasma membrane-permeable malonate ([[MitoKit-CII/Malonate-nv]]). Moreover, the SUIT-003 O2 ce D061 protocol allows evaluating the [[coupling control state |coupling control]] of living cells. Respiratory capacities are tested in a sequence of coupling states: [[ROUTINE]] and [[Electron transfer pathway]]. Optionally, to study [[LEAK respiration]], the [[phosphorylation system]] might be inhibited by [[oligomycin]]. The final concentration of [[oligomycin]] has to be carefully optimized for various cell types, to minimize the inhibitory effect on the [[Electron transfer pathway|electron transfer system]] which would lead to an underestimation of [[ET capacity]]. | |||
__TOC__ | |||
Communicated by [[Garcia-Souza LF]], [[Cardoso LHD]] and [[Gnaiger E]](last update 2019-07-02) | |||
{{Template:SUIT-003 O2 ce D061}} | {{Template:SUIT-003 O2 ce D061}} | ||
== Strengths and limitations == | == Strengths and limitations == | ||
:::+ This protocol is a control for [[SUIT-003 O2 ce D060]], with titrations of the carrier DMSO. | |||
::: | :::+ Reasonable duration of the experiment. | ||
::: | :::- To obtain [[LEAK respiration]], the concentration of [[oligomycin]] has to be carefully tested to avoid inhibitory effects in the evaluation of the [[ET capacity]]. | ||
:::- | |||
:::- Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler. | :::- Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler. | ||
== Compare SUIT protocols == | == Compare SUIT protocols == | ||
::::* [[SUIT-003 O2 ce D060]] CCP with | ::::* [[SUIT-003 O2 ce D060]] CCP with plasma membrane-permeable compounds [[MitoKit-CII/Succinate-nv]] and [[MitoKit-CII/Malonate-nv]]. | ||
::::* [[SUIT-003 O2 ce D028]] for cell membrane permeability test CCP(S). | ::::* [[SUIT-003 O2 ce D028]] for cell membrane permeability test CCP(S). | ||
::::* [[SUIT-003 O2 ce-pce D013]] first version of the Coupling control protocol and respirometric cell viability test (CCV protocol). | ::::* [[SUIT-003 O2 ce-pce D013]] first version of the Coupling-control protocol and respirometric cell viability test (CCV protocol). | ||
::::* [[SUIT-003 O2 ce-pce D018]] second version of the Coupling control protocol and respirometric cell viability test (CCV protocol). | ::::* [[SUIT-003 O2 ce-pce D018]] second version of the Coupling-control protocol and respirometric cell viability test (CCV protocol). | ||
::::* [[SUIT-003 Ce1;ce1P;ce3U;ce4Glc;ce5M;ce6Rot;ce7S;1Dig;1c;2Ama;3AsTm;4Azd]] coupling control protocol and respirometric cell viability test (CCV protocol) without oligomycin. | ::::* [[SUIT-003 Ce1;ce1P;ce3U;ce4Glc;ce5M;ce6Rot;ce7S;1Dig;1c;2Ama;3AsTm;4Azd]] coupling-control protocol and respirometric cell viability test (CCV protocol) without oligomycin. | ||
::::* [[SUIT-003 Ce1;ce2U-]] | ::::* [[SUIT-003 Ce1;ce2U-]] | ||
::::* [[SUIT-003 Ce1;ce2SD;ce3Omy;ce4U-]] | ::::* [[SUIT-003 Ce1;ce2SD;ce3Omy;ce4U-]] | ||
::::* [[SUIT-003 Ce1;ce2SD;ce3U;ce4Rot;ce5Ama]] | ::::* [[SUIT-003 Ce1;ce2SD;ce3U;ce4Rot;ce5Ama]] | ||
::::* [[SUIT-003 Ce1;ce2U;ce3Rot;ce4S;ce5Ama]] | ::::* [[SUIT-003 Ce1;ce2U;ce3Rot;ce4S;ce5Ama]] | ||
::::* [[SUIT-006]]: Coupling control protocol for mt-preparations (mtprep), counterpart to SUIT-003 for non-permeabilized cells (ce). | ::::* [[SUIT-006]]: Coupling-control protocol for mt-preparations (mtprep), counterpart to SUIT-003 for non-permeabilized cells (ce). | ||
Latest revision as of 12:36, 25 November 2020
Description
Abbreviation: CCP-ce Snv,Mnanv - control
Reference: B Carrier control for the coupling-control protocol for living cells with cell permeable compounds MitoKit-CII/Succinate-nv and MitoKit-CII/Malonate-nv - SUIT-003 O2 ce D060
SUIT number: D061_ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama
O2k-Application: O2
- MitoPedia: SUIT - Coupling-control protocol (CCP) with the carrier of the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv) and plasma membrane-permeable malonate (MitoKit-CII/Malonate-nv) in living cells (ce): DMSO.
- SUIT protocol pattern: ce1;(ce2Omy);ce3U;ce4DMSO;ce5DMSO;ce6Rot;ce7Ama
SUIT-003 O2 ce D061 protocol is a control for the SUIT-003 O2 ce D060, with the addition of the carrier DMSO instead of the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv) and plasma membrane-permeable malonate (MitoKit-CII/Malonate-nv). Moreover, the SUIT-003 O2 ce D061 protocol allows evaluating the coupling control of living cells. Respiratory capacities are tested in a sequence of coupling states: ROUTINE and Electron transfer pathway. Optionally, to study LEAK respiration, the phosphorylation system might be inhibited by oligomycin. The final concentration of oligomycin has to be carefully optimized for various cell types, to minimize the inhibitory effect on the electron transfer system which would lead to an underestimation of ET capacity. SUIT-003 O2 ce D061 protocol is a control for the SUIT-003 O2 ce D060, with the addition of the carrier DMSO instead of the plasma membrane-permeable succinate ([[[MitoKit-CII/Succinate-nv]]) and plasma membrane-permeable malonate (MitoKit-CII/Malonate-nv). Moreover, the SUIT-003 O2 ce D061 protocol allows evaluating the coupling control of living cells. Respiratory capacities are tested in a sequence of coupling states: ROUTINE and Electron transfer pathway. Optionally, to study LEAK respiration, the phosphorylation system might be inhibited by oligomycin. The final concentration of oligomycin has to be carefully optimized for various cell types, to minimize the inhibitory effect on the electron transfer system which would lead to an underestimation of ET capacity.
Communicated by Garcia-Souza LF, Cardoso LHD and Gnaiger E(last update 2019-07-02)
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
(ce2Omy) | L(Omy) | ce1;(ce2Omy)
| ||
ce3U | E | cce1;(ce2Omy);ce3U
| ||
ce4Rot | ROX | ce1;(ce2Omy);ce3U;ce4Rot
| ||
ce5DMSO | ROX | ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO
| ||
ce6DMSO | ROX | ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO
| ||
ce7Ama | ROX | ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama
|
Strengths and limitations
- + This protocol is a control for SUIT-003 O2 ce D060, with titrations of the carrier DMSO.
- + Reasonable duration of the experiment.
- - To obtain LEAK respiration, the concentration of oligomycin has to be carefully tested to avoid inhibitory effects in the evaluation of the ET capacity.
- - Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.
Compare SUIT protocols
- SUIT-003 O2 ce D060 CCP with plasma membrane-permeable compounds MitoKit-CII/Succinate-nv and MitoKit-CII/Malonate-nv.
- SUIT-003 O2 ce D028 for cell membrane permeability test CCP(S).
- SUIT-003 O2 ce-pce D013 first version of the Coupling-control protocol and respirometric cell viability test (CCV protocol).
- SUIT-003 O2 ce-pce D018 second version of the Coupling-control protocol and respirometric cell viability test (CCV protocol).
- SUIT-003 Ce1;ce1P;ce3U;ce4Glc;ce5M;ce6Rot;ce7S;1Dig;1c;2Ama;3AsTm;4Azd coupling-control protocol and respirometric cell viability test (CCV protocol) without oligomycin.
- SUIT-003 Ce1;ce2U-
- SUIT-003 Ce1;ce2SD;ce3Omy;ce4U-
- SUIT-003 Ce1;ce2SD;ce3U;ce4Rot;ce5Ama
- SUIT-003 Ce1;ce2U;ce3Rot;ce4S;ce5Ama
- SUIT-006: Coupling-control protocol for mt-preparations (mtprep), counterpart to SUIT-003 for non-permeabilized cells (ce).
References
MitoPedia concepts: SUIT protocol, SUIT B, Find
MitoPedia methods:
Respirometry