Hi,
I want to perform the HMG-CoA assay to determinate its activity in the presence of a new extract, I don't know why I think it should be used in some cell line, but so far, any protocol mentions the cells.
Can anybody explain me if this experiment is similar to DPPH assay, which doesn't use any cell line, only the extract?
Thank you in advance.
As the downstream biochemistry (e.g. the isoprenylation of small monomeric G-proteins) is quite essential, despite the clinical parameter of interest usually is cholesterol biosynthesis, I'd expect that the choice of a suitable cell line is mostly dependent on practical considerations (growth requirements, growth rate, availability, etc.). For expression levels, see e.g. here: https://en.wikipedia.org/wiki/HMG-CoA_reductase.
The advantage of using a cell extract is that you will discover potential leads which themselves do not pass an intact cell membrane, but it might not be that easy as HMG-CoA reductase is membrane bound, IIRC.
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