How to synchronize HEK293 - (Apr/14/2009 )
i'm trying to investigate subcellular localisation of certain proteins during cell cycle - it is just a pilot experiment - I would like to synchronise cells and then take pictures in time steps. At that step I don't care about having cells in a certain phase - I just want to take a look if there are any cganges during the cycle.
Does anyone know a simple method to synchronize HEK293? I don't want to buy new reagents- I was thinking about serum starvation or mitotic shake off. Does anyone have a good protocol, or a good tip how to deal with it? Let me add that I need to tranfect my cells with plasmid - when should I do it - before or after synchronisation?
I would be more than grateful.
serum starvation is the easiest: Just add serum-free medium (DMEM will be fine) for 48 hours, and then perform your experiment.