Difference between revisions of "Talk:MiPNet17.08 IOC68 Schroecken AT"
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* mouse heart homogenate prepared with PBI Shredder (position 1 for 10 seconds, position 2 for 5 seconds) and the homogenate was completely washed out of the Shredder tube with 4 ml MiR05 | * mouse heart homogenate prepared with PBI Shredder (position 1 for 10 seconds, position 2 for 5 seconds) and the homogenate was completely washed out of the Shredder tube with 4 ml MiR05 | ||
* 2.3 mg tissue per chamber = 1.15 mg / ml | * 2.3 mg tissue per chamber = 1.15 mg / ml | ||
* respiration medium: MiR05 | * Flux derivation = 20 | ||
* '''SUIT-Protocol''': PMG + AmR + HRP + | * oxygen regime 320 to 160 (reoxygenation with pure oxygen at open chamber position) | ||
* respiration medium: MiR05 (solubility factor = 0.92) | |||
* '''SUIT-Protocol''': PMG + AmR + HRP + H<sub>2</sub>O<sub>2</sub> + H<sub>2</sub>O<sub>2</sub> + D2 + D3 + S + FCCP + Rot + H<sub>2</sub>O<sub>2</sub> + H<sub>2</sub>O<sub>2</sub> + Mna + Ama | |||
* the homogenate was added, the oxygen concentration was increased till 320 nmol/ml and then the chamber was closed | |||
=== Notes === | |||
* there is a dependence of oxygen with homogenate but not that strong as with fibres (decrease of oxygen flux with decrease of oxygen concentration) | |||
* stimulation of respiration with ADP and succinate | |||
* reoxygenation with pure oxygen causes an increase in peroxide production | |||
* no big effect of FCCP on oxygen flux but a decrease in peroxide production occurred | |||
* oxygen dependence visible in the decline of oxygen flux and NO signal | |||
* rotenone increased the peroxide production | |||
* analysis of the experiments is difficult because of the O<sub>2</sub><sub>Subscript text</sub> dependence | |||
Revision as of 16:29, 23 May 2012
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IOC68 Workshop Experiments
2012-04-14 Demo Experiment
- Oxygraph AB (in both chambers the same SUIT protocol was applied)
- DatLab File 2012-04-14_AB-02
- mouse heart homogenate prepared with PBI Shredder (position 1 for 10 seconds, position 2 for 5 seconds) and the homogenate was completely washed out of the Shredder tube with 4 ml MiR05
- 2.3 mg tissue per chamber = 1.15 mg / ml
- Flux derivation = 20
- oxygen regime 320 to 160 (reoxygenation with pure oxygen at open chamber position)
- respiration medium: MiR05 (solubility factor = 0.92)
- SUIT-Protocol: PMG + AmR + HRP + H2O2 + H2O2 + D2 + D3 + S + FCCP + Rot + H2O2 + H2O2 + Mna + Ama
- the homogenate was added, the oxygen concentration was increased till 320 nmol/ml and then the chamber was closed
Notes
- there is a dependence of oxygen with homogenate but not that strong as with fibres (decrease of oxygen flux with decrease of oxygen concentration)
- stimulation of respiration with ADP and succinate
- reoxygenation with pure oxygen causes an increase in peroxide production
- no big effect of FCCP on oxygen flux but a decrease in peroxide production occurred
- oxygen dependence visible in the decline of oxygen flux and NO signal
- rotenone increased the peroxide production
- analysis of the experiments is difficult because of the O2Subscript text dependence
