Serum starvation of cells - (Nov/04/2005 )
Hi,
I'd like to know why serum starvation is used and what are the physiological condicions if I want to observe the effect of free fatty acids or glucose on the insuline signalling. Shall I use any pre-incubation in serum-free medium? and how long? the first I used 2 hours, then 12 hours without serum, but now I don't pre-incubate them and when the cells (HepG2) are grown, medium with 10% serum is replaced by medium 1%FBS and complex of fatty acids..... 
when shall I use serum starvation?
Thanks. Kofi
I'd like to know why serum starvation is used and what are the physiological condicions if I want to observe the effect of free fatty acids or glucose on the insuline signalling. Shall I use any pre-incubation in serum-free medium? and how long? the first I used 2 hours, then 12 hours without serum, but now I don't pre-incubate them and when the cells (HepG2) are grown, medium with 10% serum is replaced by medium 1%FBS and complex of fatty acids.....
when shall I use serum starvation? Thanks. Kofi
I use serum starvation in the past for metabollic labelling to make sure that the cells were in serum free medium and also to try to syncronize them as for the insulin stuff I'm not competent sorry !
pesji
Hi,
Yes seru starvation is bascially for synchronixzation of cells to the g0/g1 phase. In your case since you are looking at the insulin signaling...i suppose u will be looking at the IBP3s and IGFR etc....I think in those cases the serum starvation is basically done so that you get rid of the insulin and other growth hormones that come in the serum of whichever animal u r using...
U can also collect the media that u r conducting ur experiments in and coduct what is called a Western-Ligand blot...For more info look at articles By Cohick W (she is a prof in the animal sciences dept at rutgers and she does work on insulin and the insulin pathway)
Good luck.