Tumor cells were seeded in 96-well plates at a density of 1000–3000/well and allowed to adhere overnight. Cells were then exposed to appropriate concentrations of therapeutic agents (or vehicle control) with continuous exposure for 72 h. Growth inhibition was measured by CellTiter Aqueous MTS reagent from Promega by comparing the absorbance at 490 nm of drug-treated cells to that of untreated controls set at 100%. IC50 values were calculated using non-linear regression model (logarithmic inhibitor versus normalized response-variable slope) in Graphpad Prism 7.
Celltiter aqueous mts reagent
Manufactured by Promega
The CellTiter Aqueous MTS reagent is a colorimetric assay used to determine the number of viable cells in cell proliferation or cytotoxicity assays. The reagent contains a tetrazolium compound (MTS) and an electron coupling reagent (PES) that generate a colored formazan product when reduced by viable cells. The amount of formazan produced is proportional to the number of living cells in the sample, and can be measured spectrophotometrically.
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4 protocols using celltiter aqueous mts reagent
1
MTS Assay for Cell Viability
2
Cell Viability Assay with MTS
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Viability was assessed with the CellTiter-Aqueous MTS reagent (Promega), 24 and 48 h post-treatment. Briefly, MTS reagent was added into each well and incubated for 3h with 5% CO2 at 37°C. MTS absorbance was then measured at 490nm. Results are reported as the percentage of treated cells relative to the cells without treatment (% Control).
3
MTS Assay for Cell Viability
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Tumor cells were seeded in 96-well plates at a density of 1000–3000/well and allowed to adhere overnight. Cells were then exposed to appropriate concentrations of therapeutic agents (or vehicle control) with continuous exposure for 72 h. Growth inhibition was measured by CellTiter Aqueous MTS reagent from Promega by comparing the absorbance at 490 nm of drug-treated cells to that of untreated controls set at 100%. IC50 values were calculated using non-linear regression model (logarithmic inhibitor versus normalized response-variable slope) in Graphpad Prism 7.
4
Cell Viability Assay with MTS
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Cell viability was measured using the CellTiter Aqueous MTS reagent (Promega) according to the manufacturer’s protocol. Following optimization and accounting for growth rate differences, 1000 NCOA4-RET cells and 2000 vector, RETamp and ΔRET cells were allowed to attach per well in 96-well plates overnight for NIH/3T3 cells. For MCF10A, all cells were plated at 5000 per well. Cells were then treated with increasing concentration of inhibitors, cabozantinib or sorafenib (Selleckchem) for 72 h. After incubating with MTS reagent for 2–4 h at 37 °C, absorbance was measured at 490 nm on a plate reader (Infinite 200 PRO, Tecan). Viability of cells was calculated by dividing the average absorbance of treatment wells by that of vehicle DMSO-treated wells. Data were analyzed and represented using GraphPad Prism version 7 software.
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