Talk:Zhang 2012 Nat Protoc: Difference between revisions
From Bioblast
No edit summary |
No edit summary |
||
| Line 2: | Line 2: | ||
'''Discussion on limitations of this study:''' | '''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. | * 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 | * Short periods of high oxygen concentrations can influence stem cell properties. How did the authors transfer the stem cells from the incubator to the respirometer - was there a hypoxic atmosphere in the respirometer? | ||
* The authors | * The authors discuss the classical view on cell permeabilization and measurement of citrate synthase/lactate dehydrogenase, but do not mention the latest publications, e.g. up-to-date simple titration schemes. | ||
* | * 50 µM ADP was used. In isolated rat liver mitochondria the apparent ''K''<sub>m</sub> is 52 µM, corresponding to 50 % of OXPHOS capacity. The required ADP concentration needed to evaluate ''J''<sub>max</sub> should be minimum 9 times the ''K''<sub>m</sub> ([[Gnaiger 2001 Respir Physiol]]). | ||
Latest revision as of 07:10, 2 May 2022
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 transfer the stem cells from the incubator to the respirometer - was there a hypoxic atmosphere in the respirometer?
- The authors discuss the classical view on cell permeabilization and measurement of citrate synthase/lactate dehydrogenase, but do not mention the latest publications, e.g. up-to-date simple titration schemes.
- 50 µM ADP was used. In isolated rat liver mitochondria the apparent Km is 52 µM, corresponding to 50 % of OXPHOS capacity. The required ADP concentration needed to evaluate Jmax should be minimum 9 times the Km (Gnaiger 2001 Respir Physiol).
