6 replies to this topic

[Tabaluga]

Hi,
I would like to use the Cell Titer Glo assay (Promega), which measures ATP levels, for comparing the proliferation of several cell lines. My question is how I can do this ? Can I just measure ATP levels at one time point and conclude from the ATP level which lines proliferate more quickly ? Or do I have to measure several time points for this ? (I know it's actually a viability assay, and the method is not very exact, but still....)

Thanks !

[Curtis]

several time points. doubling time is usually around 24 hr (at least for HeLa), so I suggest you do this every 12 hr.

[Tabaluga]

Thanks for the reply. So I measure ATP levels at various time points and I can conclude that the cell lines with a higher increase of ATP over time are the ones more metabolically active i.e. faster proliferating, if I got it correctly. But does this method actually give any advantage over simply plating cells and counting them after several time points ?

[bob1]

Well, manual counting gives you about 15% error, but, in theory at least, these other assay types should give you a lower error.

[Tabaluga]

OK. I guess I also need to make a "standard curve" for each line first where I measure different cell numbers of the same line so I can later correlate lunminescence with cell number, right ?
Thanks for you reply.

[bob1]

Yes, that should work.

[Tabaluga]

Ok thanks !