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Determine cell proliferation by viability assay ?

Cell Titer Glo proliferation

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6 replies to this topic

#1 Tabaluga

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Posted 30 January 2013 - 01:51 PM

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 !

Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 


#2 Curtis

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Posted 30 January 2013 - 06:50 PM

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

#3 Tabaluga

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Posted 31 January 2013 - 04:24 AM

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 ?

Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 


#4 bob1

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Posted 31 January 2013 - 12:04 PM

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

#5 Tabaluga

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Posted 01 February 2013 - 08:04 AM

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.

Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 


#6 bob1

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Posted 01 February 2013 - 02:44 PM

Yes, that should work.

#7 Tabaluga

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Posted 01 February 2013 - 03:20 PM

Ok thanks !

Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 






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