I am using stimulated primary hepatocytes to detect alterations in AMPK activity. However, a paper published that serum starvation can increase AMPK activity in different cell types. Although, serum starvation is a standard technique used when detecting phosphorylation, I am not sure it is the best technique when trying to demonstrate differences in AMPK activation as a result of my treatments.
It depends on your purpose. Acute glucose deprivation or serum starvation can increase pAMPKa but it may last for a limited time. If you want to see the basal pAMPK level between two treatments or two phenotypes you probably do not need to starve the cells. I will not bother to use mRNA since it is a post-translational change with acute stress. If you really want to see more dramatical change in pAMPKa then to starve the cells may give you a better chance.
If you want a positive control, you can treat your cells with HBSS/HEPSS (total starvation) for 2 hours. Or with H2O2 (200 microM) for 1 hour.
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