Talk:Zhang 2012 Nat Protoc: Difference between revisions
From Bioblast
(Blanked the page) |
No edit summary |
||
| Line 1: | Line 1: | ||
'''Discussion on limitations of this study:''' | |||
* The authors use a ROCK inhibitor to prevent cells from clumping, but there is no information on whether a possible effect of this inhibitor on mitochondrial respiration has been tested. | |||
* Short periods of high oxygen concentrations can influence stem cell properties. How did the authors transport the stem cells from the incubator to the respirometer - is there hypoxic atmosphere in the respirometer? | |||
* The authors nicely discuss the classical view on cell permeabilization and measurement of Citrate Synthase/Lactate Dehydrogenase, but do not mention the latest publications. E.g. nowadays titration schemes are much simpler. | |||
* 50µM ADP was used. In isolated rat liver mitochondria the apparent K<sub>m</sub> is 52µM, corresponding to 50% of OXPHOS capacity. The required ADP concentration needed to evaluate V<sub>max</sub> should be minimum 9 times the K<sub>m</sub>. | |||
Revision as of 19:16, 2 July 2012
Discussion on limitations of this study:
- The authors use a ROCK inhibitor to prevent cells from clumping, but there is no information on whether a possible effect of this inhibitor on mitochondrial respiration has been tested.
- Short periods of high oxygen concentrations can influence stem cell properties. How did the authors transport the stem cells from the incubator to the respirometer - is there hypoxic atmosphere in the respirometer?
- The authors nicely discuss the classical view on cell permeabilization and measurement of Citrate Synthase/Lactate Dehydrogenase, but do not mention the latest publications. E.g. nowadays titration schemes are much simpler.
- 50µM ADP was used. In isolated rat liver mitochondria the apparent Km is 52µM, corresponding to 50% of OXPHOS capacity. The required ADP concentration needed to evaluate Vmax should be minimum 9 times the Km.
