I'm been trying to measure mitochondrial respiration of the cell line H9c2. I think i been having problem with the permeabilization. I wish to do de analisis using a Clark electrode. Thank you
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I send you the paper which describes how to measure cellular respiration in H9c2. If you need more details you can contact me
If you have an access to Seahorse Bioscience Flux analyzer, you can also try to measure respiration that way. Since H9C2 cells grow in a dish, they should be easily plated onto Seahorse plate. Simple protocols can be used without permeabilization, but permeabilization is also possible, e.g. with saponin.
Do you have a fluorescent plate reader? If you do check out the Luxcel Biosciences MitoXpress probes. They work very well for most lines and you can use intact cells. Permeabilision and then feed in specific mito substrates is possible.
In lieu of Seahorse, we use to use simply pulse ox or Clarke electrode measures. Its inexensive to set up (~$300) and has been around since the 1950s. No fancy kits, dyes or other tools needed. www.enzyme.chem.msu.ru/eduproc/prac/.../Oxygen%20Analyzers.doc
To study the cellular respiration you can use electron paramagnetic resonance (EPR) oximetric technique. For mitochondrial respiration you can use Seahorse Bioscience Flux Analyser technique/intrument.
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