The growth inhibitory effect of NOSH-aspirin was measured using a colorimetric MTT assay kit (Roche, Indianapolis, IN). Briefly, cells were plated in 96 well plates at a density of 104 cells per well and, following overnight incubation, were washed and treated with vehicle, NOSH-aspirin or ASA. At indicated time periods of treatment, viable cells were incubated with MTT substrate (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide, 5 mg/mL in phosphate buffered saline), further for 2 hrs, and formazan formed by viable cells was dissolved in 100 μL of the solubilization solution (10% SDS in 0.01 M HCl) and absorbance was measured on a spectrophotometric plate reader at a wavelength of 570 nm. Each experiment was performed in triplicate, and the entire experiment was repeated three times. In some experiments pan-caspase inhibitor z-VAD-FMK, pretreatment was performed prior to NOSH-aspirin addition to cells, followed by MTT assay.
Colorimetric mtt assay kit
Manufactured by Roche
The Colorimetric MTT assay kit is a laboratory equipment product from Roche. It is used to measure the metabolic activity of cells, which can be an indicator of cell viability, proliferation, or cytotoxicity. The kit utilizes the tetrazolium salt MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) as the core reagent.
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4 protocols using colorimetric mtt assay kit
1
MTT Assay for NOSH-Aspirin Cytotoxicity
2
Antiproliferative Effects of NOSH-ASA Isomers
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Cell growth inhibitory effect of the positional isomers of NOSH-aspirin was measured using a colorimetric MTT assay kit (Roche, Indianapolis, IN).
HT-29 and HCT-15 cancer cells were plated in 96-well plates at a density of 30,000 cells/well. The cells were incubated for 24 h with different concentrations of positional isomers of NOSH-ASA. The absorbance of the plates was measured on a spectrophotometric plate reader at a wavelength of 570 nm.
HT-29 and HCT-15 cancer cells were plated in 96-well plates at a density of 30,000 cells/well. The cells were incubated for 24 h with different concentrations of positional isomers of NOSH-ASA. The absorbance of the plates was measured on a spectrophotometric plate reader at a wavelength of 570 nm.
3
Colorimetric Cell Viability Assay
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Cell metabolic activity was determined by the colorimetric MTT assay kit (Roche Diagnostics, SL) in which tetrazolium salts (MTT) are reduced to formazan by the mitochondrial succinate-reductase system. Cells were seeded in 96-well microplates and after treatment were incubated with 10 μl of MTT labelling reagent for 4 h, and then the formazan dye formed was dissolved with 100 μl of solubilisation solution. The absorbance was measured using a multiwell spectrophotometer, to estimate the number of viable cells.
4
Cell Proliferation and Cytotoxicity Assay
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Cell proliferation viability and cytotoxicity assay were performed as described previously [13 (link)]. Cells were seeded, and then the colorimetric MTT Assay kit (Roche Diagnostics) was used to measure the cell proliferation until 72 hours. For the foci formation assay, cells were seeded, cultured, fixed, and stained.
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