SUIT-003 AmR ce D059: Difference between revisions
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::* It is recommended to run this test with every different sample type used. | ::* It is recommended to run this test with every different sample type used. | ||
::+ Reasonable duration of the experiment. | ::+ Reasonable duration of the experiment. | ||
::- The concentration of [[oligomycin]] has to be carefully tested to avoid inhibitory effects in the evaluation of the [[ET | ::- 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. | ||
Revision as of 16:06, 8 June 2020
Description
Abbreviation: Amr effect on ce - control
Reference: B: Experimental control for SUIT-003 AmR ce D058
SUIT number: D059_ce1;ce1H2O;ce1MiR05;ce1DMSO;ce2Omy;ce3U;ce4Rot;ce5Ama
O2k-Application: AmR
- MitoPedia: SUIT - Carrier control protocol to SUIT-003 AmR ce D058
- SUIT protocol pattern: ce1;ce1H2O;ce1MiR05;ce1DMSO;ce2Omy;ce3U;ce4Rot;ce5Ama
SUIT-003 AmR ce D059 is a control protocol of SUIT-003 AmR ce D058 to study the effect of the carrier titrations on the respiration in a parallel experiment. This protocol should be run in parallel with SUIT-003 AmR ce D058 to compare the respiratory capacities and the Flux Control Ratios (FCR) in order to detect any inhibition of the respiratory capacities by the Amplex UltraRed or other components of the ROS assay.
Communicated by Garcia-Souza LF, Komlodi T and Gnaiger E(last update 2019-06-26)
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
ce1H2O | ROUTINE | ce1;ce1H2O
| ||
ce1MiR05 | ROUTINE | ce1;ce1H2O;ce1MiR05
| ||
ce1DMSO | ROUTINE | ce1;ce1H2O;ce1MiR05;ce1DMSO
| ||
ce2Omy | LOmy | ce1;ce1H2O;ce1MiR05;ce1DMSO;ce2Omy
| ||
ce3U | E | ce1;ce1H2O;ce1MiR05;ce1DMSO;ce2Omy;ce3U
| ||
ce4Rot | ROX | ce1;ce1H2O;ce1MiR05;ce1DMSO;ce2Omy;ce3U;ce4Rot
| ||
ce5Ama | ROX | ce1;ce1H2O;ce1MiR05;ce1DMSO;ce2Omy;ce3U;ce4Rot;ce5Ama
|
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Coupling control
- Pathway control
- ยป Electron transfer pathway
- ยป Fatty acid oxidation pathway control state, F
- ยป NADH electron transfer-pathway state, N
- ยป Succinate pathway control state, S
- ยป NS-pathway control state, NS
- ยป Glycerophosphate pathway control state, Gp
- ยป Complex IV single step, CIV
- ยป Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- ยป Glutamate, G
- ยป Glycerophosphate, Gp
- ยป Malate, M
- ยป Octanoylcarnitine, Oct
- ยป Pyruvate, P
- ยป Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- This protocol has been designed to study the effect of the carrier titrations on the respiration in parallel to SUIT-003 AmR ce D058. Therefore, instead of SOD, HRP and AmR, the carriers H2O; MiR05; and DMSO are titrated.
- It is recommended to run this test with every different sample type used.
- + Reasonable duration of the experiment.
- - 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 AmR ce D058: is a protocol to study the inhibitory effect of the AmR assay towards the respiration
- SUIT-003 O2 ce D009 for non-permeabilized cells with oligomycin without the AmR assay
References
MitoPedia concepts: SUIT protocol, SUIT B, Find
MitoPedia methods:
Respirometry