Usually it is mentioned that incubation for 1-4 hours with MTS reagent before reading. I have tried cancer cell line(incubated for72 hours with drug) for 1 hour, 2 hour, and 3 hour incubation with MTS reagent. The viability % is different according to time. So, which data should I believe on..1 hr,2 hr or 3 hr?
How to determine the MTS reagent incubation time for different cell lines?
Thank You
Kamal Pandey
Actually, I think both time of incubation and cell concentration matters in an MTS assay. A long time incubation results in an over consumption of MTS and have not difference of each treatment, for cells in each well all reach the platform stage. In my experience,while dealing with tumor cells, we plant the cell at a concentration from 3*103 to 1*104 per well and incubate MTS for 2 hrs to prevent over reaction.
Hope this might help. Regards.
All cells have different ability to metabolize the MTS. So some cells need more time than others to show the results.
In addition the confluence in which the cells are also is a point to take into account. At least 50% confluency is necessary, especially if the cells metabolize little.
You should adjust the number of cells so that in 2 or 3 hours at most you can have an absorbance between 0.8 and 1.2 and then under those conditions perform your experiment.
hope this might help
regards
In human panctratic beta cell lines (1.1B4), i have incubated 100000 cells per well with MTS reagent for one hour and i got good response.
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