fatty acids-induced HepG2 cells and how prepare the extract in DMEM medium
Dear Sylvestar Darvin,
Free fatty acids-induced HepG2 cells?
I did not understand the question. Free fatty acid induced what in HepG2 cells?
HepG2 cells grown in DMEM with low glucose dont go for high glucose.
I think you want to induce apoptosis in HepG2 cells by fatty acids and you want to see the protective effect of plant extracts you are working with right?
Dissolve your plant extract in DMSO and pre-incubate with cells in 96 well plate, keep both positive and negative control.
Do MTT assay to confirm the protective effect of the plant extract.
If your plant extract is effective cells dont die other wise it will.
sir my work is I induced lipid with sodium oliate + plamatic acid + bsa (fatty acid conjugation) in to the cells and after that i treat with compound and see reduction in lipids so i need to prepare the that Fatty acid conjugation and need to dissolved my compund in DMSO and media need to fix concentration ext.. pleas help
I have little experience in that assay. however, if you want to induce apoptosis then you will need very high concentration of FFA (>1.0mM according to some refs.) I think measuring the decrease of lipid accumulation would be more general protocol.
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