Viability was measured using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. MIA-PaCa2 or MIA-PaCa2-GEM Cells were plated in 96-well plates overnight before the gemcitabine or vehicle treatment. For siRNA or miR mimic/inhibitor treatment, the cells were transfected with RNA Oligoribonucleotides in the presence or absence of 100 nM gemcitabine. At the end of the treatment, 10% v/v of 5 mg/ml solution of MTT agent (Sigma-Aldrich) was added for 2 h. The medium was then removed and the cells were dissolved in DMSO (Sigma-Aldrich). Relative cytotoxicity was determined by measuring the absorbance at 570 nm using a luminometer (Molecular Devices, U.S.A.).
Mtt agent
Manufactured by Merck Group
Sourced in United States
MTT agent is a colorimetric assay used to measure cell metabolic activity. It is a commonly used reagent for quantifying viable cells in proliferation or cytotoxicity assays.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Xmark microplate absorbance spectrophotometer, by Bio-Rad (1 mentions)
Rpmi 1640 medium, by Wuhan Boster Biological Technology (1 mentions)
Multiskan spectrum microplate reader, by Thermo Fisher Scientific (1 mentions)
Elx800, by Agilent Technologies (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Ly294002, by Beyotime (1 mentions)
14 protocols using mtt agent
1
Cytotoxicity Assay for Pancreatic Cancer
2
Cell Proliferation Measurement via MTT Assay
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MTT assay was performed to detect cell proliferation. Following treatment, the cells were inoculated in a 96-well plate with 1×104 cells per well and incubated in a 37°C, 5% CO2 incubator. Then, 20 µl 5 mg/ml MTT agent (Sigma-Aldrich; Merck KGaA) was added to cells. Following incubation for 4 h, the formazan crystals were dissolved in 150 µl DMSO (Sigma-Aldrich; Merck KGaA). The absorbance was measured at a wavelength of 490 nm using a microplate reader.
3
Evaluating 5-Azacytidine's Cytotoxicity in Cells
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The cell lines were seeded in 96-well plates at a concentration of ~3×104 cells/mL. When the cells formed a confluent monolayer, they were treated with 5-azacytidine (0, 0.25, 0.5, 1, 2.5, 5, 10, 25, 50, and 100 μM) for 24, 48, or 72 h, respectively. Next, 20 μL MTT agent (5 mg/mL; Sigma-Aldrich) was added to the cells and was incubated for 4 h at 37°C. The supernatants were then removed and 150 μL of dimethyl sulfoxide (Sigma-Aldrich) was added. The plates were shaken and incubated at 37°C for 10 min. Absorbency was measured using a microplate reader (BioTek, Vinooski, VT, USA) at 490 nm. Each experiment was repeated at least five times. Inhibition of cell growth was calculated using the following formula:
4
MTT Assay for Cell Viability
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The cells were seeded in 24 well plates and incubated with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT agent (Sigma-Aldrich, M-2128) diluted at 0.5 mg/ml for 3 h. The plates were then covered and kept in the incubator at 37 °C to allow reduction of the yellow tetrazolium salt to purple formazan crystals by metabolically active cells. The insoluble crystals were dissolved by using the isopropanol 0.4% HCl and the optic density was measured at 550 nm using the Bio-Rad xMark™ Microplate Absorbance Spectrophotometer. IC50 is defined as the half maximal inhibitory concentration required to inhibit a biological process by half. Three technical replicates and four biological replicates were carried out.
5
Evaluating liposIA Cytotoxicity on Cell Lines
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The cytotoxicity of liposIA on various cell lines was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Briefly, SV40, HK-2, HEK293T and C2C12 cells were seeded on 96-well plates at 1 × 104 cells per well and cultured for 24 h in a 5% CO2 humidified atmosphere and DMEM with 10% final concentration of FBS at 37°C. Then, the culture medium was refreshed with 100 μL DMEM containing different concentrations of liposIA. After 24 h of incubation, the cells were rinsed with PBS. MTT agent (Sigma, USA) was subsequently added in each well at a concentration of 0.5 mg/mL at 37°C for 4 h. Finally, 100 μL of dimethyl sulfoxide (DMSO) was employed to dissolve formazan by constant shaking, whose absorbance was determined with a microplate reader at 570 nm.
6
Bacterial Viability Assay of Canal Samples
7
Cell Viability Assays: Cell Counting and MTT
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Cell counting assay and MTT assay were used to measure the cell viability. Cell counting was performed using a cell counter. MTT assay was performed after plating 2 × 103 cells in each well and after 24 h, cells were added with 10 μL of MTT agent (Sigma, St. Louis, MO, United States) and incubated for another 4 h. Following this, cell medium was removed and 20 μL DMSO was added to dissolve the precipitates and a plate reader was used to measure the absorbance at 480 nm.
8
Assessing NSCLC Cell Viability
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The viability of NSCLC cells was measured using the MTT assay. In brief, cells that transfected with pcDNA-caspase-1 were seeded into 96-well plates at a density of 10,000 cells per well, followed by treatment with Alimta at different doses for 24 hours. Then, the MTT agent (0.5 mg/ml; Sigma, USA) was added into each well and incubated for another 4 hours. The medium was removed, and 150 μl dimethyl sulfoxide (DMSO) was added to each well to incubate for 10 minutes at room temperature. The absorbance values at 490 nm were detected using a microplate reader (Thermo, USA). For depiction of the cell growth curve, cells were transfected with indicated oligonucleotides or cotreated with Alimta at 100 nm for 0, 24, 48, and 72 hours, respectively.
9
Cell Viability Assay using MTT
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Plates with 96 wells were used to culture 2,000 cells in 100 µL of media. Every well received an additional 10 µL of MTT agent (Sigma-Aldrich Co., St Louis, MO, USA) at 5 mg/mL concentration. The media were drained after 4 hours. To promote the lysis of formazan crystals, 150 µL of DMSO was added to every well. The absorbance was recorded at 490 nm using a Thermo Fisher Spectrophotometer 1510 (Molecular Devices LLC, Sunnyvale, CA, USA). The survival rate was calculated as a ratio of the absorbance of supplemented cells to that of the control groups. This process was repeated thrice.
10
Mitochondrial-Dependent Cell Viability Assay
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Cell viability was determined by the mitochondrial-dependent reduction of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) to formazan by adding 10 μL of the MTT agent (5 mg/mL; Sigma-Aldrich) to cells in the plates [35 (link)].
LDH activity was detected using the LDH activity assay kit according to the manufacturer's instructions.
LDH activity was detected using the LDH activity assay kit according to the manufacturer's instructions.
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