Survival rate in MTT - (Mar/17/2009 )
Hi,
Has any one know how should I calculate the survival rate after getting the OD from MTT.
Many thanks,
Sara
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.
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
saraarasus on Mar 18 2009, 10:46 PM said:
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)
cotchy on Mar 20 2009, 01:24 AM said:
Wouldn't you need a higher concentration than 1% DMSO?
0.01% DMSO is sufficient to kill most cell lines, it dissolves essential cell membrane components.
bob1 on Mar 19 2009, 05:00 PM said:
Thanks for your helpful reply.
Cheers,
Sara