MTT dye reduction assay (Sigma, St. Louis, MO, USA) was carried out to detect the viability of SW1990 and BxPC-3 Cells as previously reported (Dai et al., 2012 (link)). Briefly, cells were seeded into a 96-well plate at a density of 1 × 104 cells/well, cultured for 12 h, then treated with Hellebrigenin with different concentration (0,6, 12, 24, 48, 96 nM in SW1990 and 0, 3.125, 6.25, 12.5, 25, 50 nM in BxPC-3) for 0 to 96 h. At the end of the treatment, 10 μL (50 µg) MTT solution was added into the cells and incubated for another 4 h. 200 μL Dimethylsufloxide (DMSO) was added to each well after removal of the supernatant. After shaking for 10 min, cell viability was measured at the absorbance of 490 nm using an Enzyme-labeling instrument (Multiskan. Go, Thermo Scientific, Ratastie, Finland). The experiments were repeated for three times. Cell growth curve was completed using time as the abscissa and a value (mean ± SEM) as the ordinate.
Mtt dye reduction assay
Manufactured by Merck Group
Sourced in United States
The MTT dye reduction assay is a colorimetric technique used to assess cell metabolic activity and viability. It measures the reduction of the yellow tetrazolium dye MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) to purple formazan crystals by metabolically active cells. The degree of color change is proportional to the number of viable cells present in the sample, which can be quantified using a spectrophotometer.
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6 protocols using mtt dye reduction assay
1
MTT Assay for Cell Viability in Pancreatic Cancer
2
Assessing Cell Viability with MTT Assay
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The cell viability was assessed using a standard MTT dye reduction assay, according to the manufacturer's instructions (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany). Accordingly, each of the three substances, ZINC05463076, ZINC2102846, ZINC19901103, were analyzed at five different concentrations ranging from 0.01 to 100 µmol, 6.25, 12.5, 25, 50 and 100 µmol. Briefly, 3×105 PC-3, DU-145, LNCaP or LNCaP-AI were plated into each well of 6-well flat-bottomed microplates and incubated overnight at 37°C in 5% CO2. Subsequently, they were untreated (controls) or treated for 72 h with each experimental compound at 37°C. Results were expressed as the mean cell number.
3
Cell Proliferation Assay of TP53 Mutants
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Proliferation rates of the two different TP53 mutant cell lines were investigated in comparison to the vector control cell line using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) dye reduction assay (Sigma). Briefly, 3000 cells were seeded in the wells of 96-well plates in triplicates in complete DMEM medium, and the cells were allowed to grow for 48 h. MTT reagents were added to each well and incubated for 2 h in the cell culture incubator. After 2 h, the culture media was carefully removed and 150 μL of DMSO added. The plates were incubated on a shaker for 10 min, and the absorbance was measured at 570 nm.
4
MTT Assay for Cell Viability
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Cell viability was measured by the [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) dye reduction assay (Sigma-Aldrich, St. Louis, USA). Cells were plated in 96-well plates at a density of 3 × 103 cells/100 μL/well followed by 24 h incubation. Cells were then treated with RPS at 0, 2.5, 5, 7.5, 10, 15, 20, 30, 40, or 60 μg/mL for 48 hours. After treatment, 20 μL of MTT (5 mg/mL) was added to each well and incubated for 4 hours. The media were then removed and the dark blue formazan crystals were dissolved in 150 μL of DMSO. The absorbance at 550 nm was measured in a microplate reader (Corning Costar, Corning, NY, USA). The background absorbance at 690 nm was subtracted from all readings. The percentage of cell viability relative to the controls without RPS was calculated. Each concentration was tested with 6 repeats in each experiment, and the experiment was repeated independently at least 3 times.
5
MTT Dye Reduction Assay for Cell Viability
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MTT dye reduction assay (Sigma, St.Louis, Mo, USA) was carried out to detect the cell viability as previously reported (Dai & etal, 2012 (link)). Briefly, cells were seeded into a 96-well plate at a density of 1 × 105 cells/well, cultured for 12 h, then treated with LPS at a concentration of 300 μ g/mL for 24 h, and then treated with MSC-exosomes for 24 h. At the end of the treatment, 10 μ L (50 μ g) of the MTT solution was added into the cells and incubated for another 4 h. Two hundred microliters of dimethylsufloxide (DMSO), was added to each well after removal of the supernatant. After shaking for 10 min, cell viability was measured at an absorbance wavelength of 490 nm using an Enzyme-labeling instrument (Multiskan. Go, Thermo Scientific, Ratastie, Finland).
6
Cell Viability Evaluation by MTT Assay
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Cell viability was evaluated by 3-(4,5-Dimethyl-2-thiazol)-2,5-diphenyl-2H-tetrazolium bromide (MTT) dye reduction assay according to the manufacturer protocol (Sigma Aldrich). Drug- or vehicle-treated cells were incubated with MTT dye (at final concentration of 0.5 mg/mL) and solubilized by DMSO. Absorbance was measured by a spectrophotometer at 570 nm.
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