DLD-1 cells were seeded into 96-well plates at a density of 1 × 103 cells/well and stabilized for 24 h. DLD-1 cells were then treated with IL-17C (100 ng/mL; eBioscience) for 24 h. Subsequently, 20 µL of MTT solution (5 mg/mL) (Duchefa Biochemie, Haarlem, The Netherlands) was added to each well and incubated for 4 h. The MTT solution was then removed, and 150 µL of DMSO was added to each well to dissolve the formazan crystals. Absorbance was measured at 540 nm using the Multiskan™ GO Microplate Spectrophotometer (Thermo Fisher Scientific, Fair Lawn, NJ, USA).
Mtt solution
Manufactured by Duchefa Biochemie
Sourced in Germany
MTT solution is a laboratory reagent used for cell proliferation and cell viability assays. It is a yellow tetrazolium salt that can be reduced by metabolically active cells to form purple formazan crystals. The formation of these crystals is quantifiable and can be used to measure cellular metabolic activity, which is often correlated with cell viability and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Duchefa Biochemie (2 mentions)
Synergy ht, by Agilent Technologies (1 mentions)
Mtt solution, by Merck Group (1 mentions)
1.5 ml reaction tubes, by Greiner (1 mentions)
Phosphate buffered saline solution, by Harvard Bioscience (1 mentions)
Multiskan go microplate spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Spectramax m3, by Molecular Devices (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Duchefa Biochemie (1 mentions)
7 protocols using mtt solution
1
MTT Assay for DLD-1 Cell Viability
2
Assessing Microglia and Neuron Viability
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Cell viability of BV-2 microglia and primary neurons was detected by MTT assay. In brief, 100 μl of MTT solution (0.5 mg ml−1, Duchefa) was added to cultured cells and incubated for an additional 4 h at 37°C, until the medium turned purple. Absorbance at 570 nm was measured by a microplate reader after addition of 100 μl DMSO. Each experiment was performed in triplicate and repeated three times with separate cell preparations. Results were expressed as a percentage of the control value at 570 nm.
3
MTT Assay for Cell Viability
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Cellular viability was determined using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The HEI-OC1 cells were exposed to 0.5 mg/mL of MTT solution (Duchefa Biochemie, Amsterdam, The Netherlands) for 4 h, and the resulting formazan crystals were solubilized with 100 μL of dimethyl sulfoxide (DMSO). The absorbance was measured using a 96-well microplate reader (Synergy HT, BioTek Instruments, Winooski, VT, USA) at 570 and 630 nm. The average optical density (OD) in the control cells was taken as 100% viability.
4
Cytotoxicity Assay for Cancer Cells
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A549, H460, HCC-1359, HCC-366, H2087 (1.5 × 104 cells/mL) and WI-26 cells (4 × 104 cells/mL) were seeded and cultured in 96-well flat bottom plates for 24 h and treated with chemicals in serum-free medium for 72 h. After incubation, 10 μL of MTT solution (5 mg/mL, Sigma) was added and incubated for 1.5 h at 37 °C. After the medium containing MTT solution was discarded, the precipitated formazan crystals were dissolved in 100 μL of DMSO (Duchefa) per well. The absorbance was measured at 560 nm using a microplate reader (Sunrise, TECAN, Männedorf, Switzerland). GI50 was calculated by using the Prism (GraphPad).
5
Quantifying Cell Viability via MTT Assay
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The cell vitality was determined using an MTT assay (Mosmann 1983 (link)) as described in Sachs et al. (2017 ). This approach measures the activity of mitochondrial and cytosolic dehydrogenases, which reduces the yellow, water soluble 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) to a blue, water-insoluble formazan product (Lindl and Gstraunthaler 2008 ).
After cell exposure to Eu(III) or U(VI), 50 mg of fresh cells were weighed into 1.5-mL reaction tubes (Greiner) followed by the addition of 1-mL phosphate-buffered saline solution without Ca2+ and Mg2+ (PBS; Biochrom, Berlin, Germany) and 200 μL MTT solution (5 mg/mL; Duchefa, Harlem, The Netherlands). Subsequently, the assay was performed as described in Sachs et al. (2017 ). The vitality of the Eu(III) and U(VI) exposed cells was determined as a percentage of the control samples according to Eq. (1 ).
The results represent data from five independent experiments, each with two to three samples for control and each metal concentration.
After cell exposure to Eu(III) or U(VI), 50 mg of fresh cells were weighed into 1.5-mL reaction tubes (Greiner) followed by the addition of 1-mL phosphate-buffered saline solution without Ca2+ and Mg2+ (PBS; Biochrom, Berlin, Germany) and 200 μL MTT solution (5 mg/mL; Duchefa, Harlem, The Netherlands). Subsequently, the assay was performed as described in Sachs et al. (2017 ). The vitality of the Eu(III) and U(VI) exposed cells was determined as a percentage of the control samples according to Eq. (
The results represent data from five independent experiments, each with two to three samples for control and each metal concentration.
6
Cell Viability Assay using EPF
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First, 3 × 103 cells/well were seeded into 96-well plates and stabilized overnight. These cells were treated with EPF at different concentrations (0.1–100 μg/mL) and incubated at 37 °C for 24 h. At 24 h post treatment, MTT solution (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; Duchefa, Haarlem, The Netherlands) was added to the culture medium at 0.4 mg/mL. After 2 h of incubation at 37 °C, the culture medium was discarded, and 100 μL DMSO was added into each well to thoroughly dissolve MTT formazan. Cell viability was evaluated by measuring the absorbance of each well at 540 nm using a microplate reader (SpectraMax M3; Molecular Devices, San Jose, CA, USA).
7
MTT Assay for Cell Viability
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3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT; Duchefa Biochemie, Haarlem, The Netherlands) assay was performed to determine cell viability. J774 A.1 macrophage cells were seeded in 96-well plates at a density of 1.25 × 105 cells/well and incubated for 16 h. Cells were pre-treated with KPA (3.125-100 μM) isolated from root of C. sinica for 1 h and then stimulated 1 μg/ml LPS (Sigma Aldrich, St. Louis, MO, USA) for 24 h. After incubation, the medium was removed and 100 μl MTT solution (Duchefa Biochemie, Haarlem, The Netherlands) was added to each well and the cells were more incubated for 2 h. After then the supernatant was removed and dissolved in 100 μl dimethyl sulfoxide (DMSO; Duchefa Biochemie, Haarlem, The Netherlands) in each well. Absorbance at a wavelength of 540 nm was measured using a SpectraMax 190PC microplate reader (Molecular Devices, Sunnyvale, CA, USA). The experiment data were expressed as the means ± standard deviation of triplicate assay.
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