6 replies to this topic

[saraarasus]

Hi,
Has any one know how should I calculate the survival rate after getting the OD from MTT.

Many thanks,
Sara

[WOW]

According to my understanding MTT assay in most cases reflects "cell viability". This term was somehow different from cell survival or cell proliferation. Therefore once you do this assay in a short time (within 24hrs), it mostly reflect cell death and survival. If the time is more than 24hrs, you should consider proliferation effect. Since I am not clear your experiment design, I could not exactly answer your questions. May it help you and good luck.

[saraarasus]

Hi WOW,
Thanks for your reply. What I did was that, I treated my cells for 24 hours with my protein. Then I did the MTT. I remove the media , then add 90micro litre of RPMI without phenol red, then I added 10microlitre of 5mg/ml MTT in the same media and incubate it for 4 hrs at 37 deg. Then I added MTT solvent and mix them and read the OD whithin 1 houre.
I have two question:
1-Is it the correct procedure?
2- What should I use as the high control?

Thanks,
Sara

[cotchy]

saraarasus, on Mar 18 2009, 10:46 PM, said:

Hi WOW,
Thanks for your reply. What I did was that, I treated my cells for 24 hours with my protein. Then I did the MTT. I remove the media , then add 90micro litre of RPMI without phenol red, then I added 10microlitre of 5mg/ml MTT in the same media and incubate it for 4 hrs at 37 deg. Then I added MTT solvent and mix them and read the OD whithin 1 houre.
I have two question:
1-Is it the correct procedure?
2- What should I use as the high control?

Thanks,
Sara

Sara your protocol looks fine.

A positivie control of DMSO can be used (1 % or 0.1% DMSO dissolved in your normal media is sufficent to kill cells)

[DRT]

cotchy, on Mar 20 2009, 01:24 AM, said:

A positivie control of DMSO can be used (1 % or 0.1% DMSO dissolved in your normal media is sufficent to kill cells)

Wouldn't you need a higher concentration than 1% DMSO?

[bob1]

0.01% DMSO is sufficient to kill most cell lines, it dissolves essential cell membrane components.

[saraarasus]

bob1, on Mar 19 2009, 05:00 PM, said:

0.01% DMSO is sufficient to kill most cell lines, it dissolves essential cell membrane components.

Thanks for your helpful reply.

Cheers,
Sara