HCC (Hep3B, Huh7) and NSCLC (A549, H1299) cell lines were seeded in a 96-well plate at a density of 2.5 × 104 cells/mL in complete media and allowed to adhere for 24 h. The cells were serum-starved (1% serum containing medium) for 24 h and were then treated with different concentrations (0.1, 0.3, 1, 3, 10, 30, 100 μM) of cabozantinib (Selleckchem.com, Huston, USA), compound 3 , or compound 4 in the presence of 10% serum. After 48 h, 25 µL of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) dye solution (5 mg/mL) (Merck, MA, USA) was added to the wells and further incubated for 4 h at 37 °C. Then, the media were removed, and 200 µL of dimethyl sulfoxide (DMSO) (Duksan, Ansan, South Korea) was added to solubilize the formazan crystal. After a 30 min incubation, the absorbance was measured at 540 nm using a microplate reader (BMG LABTECH, Ortenberg, Germany).
Mtt dye solution
Manufactured by Merck Group
Sourced in United States
MTT dye solution is a colorimetric assay used to measure cell metabolic activity. It is a water-soluble tetrazolium salt that is reduced by metabolically active cells, producing a colored formazan product that can be quantified using a spectrophotometer.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Duksan Pure Chemicals (2 mentions)
Microplate reader, by BMG Labtech (2 mentions)
Matrigel, by BD (2 mentions)
Elx800, by Agilent Technologies (2 mentions)
Cabozantinib, by Selleck Chemicals (1 mentions)
Enzalutamide, by Selleck Chemicals (1 mentions)
Microplate reader, by Thermo Fisher Scientific (1 mentions)
680 microplate reader, by Bio-Rad (1 mentions)
Elisa plate reader, by Agilent Technologies (1 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
N 3 dimethylaminopropyl n ethylcarbodiimide hydrochloride, by Merck Group (1 mentions)
N hydroxysuccinimide, by Merck Group (1 mentions)
3 mercaptopropionic acid, by Merck Group (1 mentions)
Putrescine, by Merck Group (1 mentions)
Lipase from thermomyces lanuginosus, by Merck Group (1 mentions)
N boc serinol, by Merck Group (1 mentions)
Polycaprolactone diol, by Merck Group (1 mentions)
Lymphoprep solution, by Axis-Shield (1 mentions)
19 protocols using mtt dye solution
1
Cytotoxicity Evaluation of Cabozantinib and Compounds
2
Cytotoxicity Assay of Anti-cancer Compounds
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LNCaP and PC-3 cells were seeded in 96-well plates at a density of 2.5×104 cells/mL in serum-starved conditions (1% FBS). After 24 h, the cell medium was changed to 10% serum-containing medium with various concentrations (0.1, 0.3, 1, 3, 10, 30, and 100 μM) of enzalutamide (Selleckchem, Houston, TX, USA), navarixin (Selleckchem), or MMP-2/9 inhibitor (Sigma-Aldrich, St. Louis, MO, USA). After 48 h, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) dye solution (5 mg/mL; Merck, Kenilworth, NJ, USA) was added to the wells and incubated for 4 h at 37°C. The media were then discarded, and 200 µL of dimethyl sulfoxide (DMSO) was added to each well (Duksan, Ansan, Korea) to solubilize formazan crystals. After 30 min of incubation, the absorbance was measured at 540 nm using a microplate reader (BMG Labtech).
3
MTT Cell Viability Assay Protocol
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Cells were seeded in each well of a 96-well plate 24 hr prior to treatment. Dimethylthiazolyl-2-5-diphenyltetrazolium bromide (MTT) dye solution (Sigma, St. Louis, MO, USA) was added into the 96-well plate 20 hr post treatment. The plate was incubated at 37°C for 4 hr, and the treatment terminated by adding stop solution (isopropanol with 0.04 N HCl). MTT was cleaved by live cells to a colored formazan product. The supernatant was discarded and 150 μL DMSO was added to each well, mixed evenly, and the absorbance (A) was measured at 450 nm within 10 min using a DG-5031 ELISA Reader [21 (link)].
4
CRC Cell Proliferation, Migration, and Invasion Assays
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First, CRC cells were seeded in 96-well plates at a density of 20,000 cells per well. After transfection as previously described and 0, 24, 36, 48, 72 or 96 h after treatment, 20 μL of MTT dye solution (5 mg/mL, Sigma, NY, USA) was added to each well. After incubation for another 4 h, 150 μL of DMSO was added to each well. The optical density (OD) was measured at 490 nm. For the invasion and migration assays, 1 × 105 cells were cultured in the upper chamber of a 24-well plate in serum-free medium, whereas the bottom chamber was supplemented with complete medium. After 24 h of culture, the migrated cells were washed, fixed, and stained with 0.1% crystal violet for 10 min. Representative images were captured under a light microscope. In addition, cell invasion was similarly detected based on the addition of inserts precoated with Matrigel (BD Biosciences, USA), which can mimic invasion-triggering circumstances. All of these assays were performed using at least three independent experiments.
5
Cell Viability and Clonogenicity Assays
6
MTT Cell Viability Assay
7
Cell Growth and Colony Formation
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Cell growth was evaluated by MTT assay. Briefly, Cells were seeded in 96-well plates (1000 cells/ well) in triplicate and cell viability was examined by MTT dye solution (5 mg/ ml, Sigma) every two days. For colony formation assay, cells were seeded in 6-well plates (800cells/ well) in triplicate and cultured under normal growth conditions for two weeks. Colonies were washed and stained with 0.1% crystal violet, and were counted using an inverted microscope.
8
Cell Viability Measurement using MTT Assay
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Cell viability was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay according to the manufacturer's instructions. Briefly, cells were seeded in 96-well plates at a density of 1 × 104 cells per well and after 36 h of growth in a humidified atmosphere at 37 °C and 5% CO2, 20 μl of MTT dye solution (5 mg ml−1, Sigma, Grand Island, NY, USA) was added to each well and incubated for 4 h at 37 °C. After the addition of dimethyl sulphoxide (150 μl), plates were incubated for an additional 10 min and absorbance was measured using a microplate spectrophotometer (ELx800, Bio-TEK, Winooski, VT, USA) at a wavelength of 570 nm.
9
Quantifying Gastric Cancer Cell Proliferation
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A modified MTT assay was used to determine the proliferation of human gastric cancer cells. Briefly, cells were seeded in 96-well plates at a density of 1×104 cells/well and treated with Tan IIA. Following 24 h culture at 37°C, 20 µl MTT dye solution (Sigma-Aldrich; Merck KGaA) was added to each well and incubated for a further 2 h. The medium was subsequently removed and replaced with Solubilization/Stop solution (100 µl/well; Sigma-Aldrich; KGaA). Absorbance was measured at 570 nm using a microplate reader (Thermo Fisher Scientific, Inc.).
10
Sorafenib Cell Viability Assay
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Cell viability was assessed by using the 3-(4,5-dimethylthiazol-2yl)-2,5- diphenyl-2H-tetrazolium bromide (MTT) assay. In brief, cells were seeded in 96-well plates (5000 cells/well) and incubated for 24 h at 37°C. Then, sorafenib was added to the wells at various concentrations, and the plates were incubated for another 72 h at 37°C. MTT dye solution (Sigma-Aldrich, St. Louis, MO) was added to the wells, and the plates were incubated at 37°C for 4 h, followed by addition of stop solution (isopropanol with 0.04 N HCl). Absorbance at 570 nm was determined on a (Bio-Rad, Hercules, CA) plate reader. IC50 values were determined by non-linear regression analysis. Values are given as mean ± standard deviation of 5 experiments.
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