In my experiments, there is an accumulation of malonyl-CoA in HepG2 cells. I need to demonstrate whether this accumulation is inhibiting beta-oxidation pathway or not.
One way is to measure fatty acid-dependent oxygen consumption in intact cells using either a Clark-type electrode or a Seahorse XF analyzer. This however can be troublesome in intact cells because you'd have to eliminate the presence of other respiratory substrates first (i.e., glucose, pyruvate, and glutamine) in order to cleanly assess how oxygen consumption changes as a function of fatty acid addition. The amount of time needed for the deprivation prior to assay is going to be cell-type dependent.
The second way is if you can use a fatty acid substrate labeled with a stable isotope to measure production acid soluble metabolites using mass spec or NMR. I'm not sure of a simpler solution at the moment.
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