Cell viability was determined by MTS Assays. HeLa cells were seeded 3000 cells in a 96-well plate and incubated overnight. Cells (2–5 × 104) were treated with EGCG (25 μM), cisplatin (250 nM), and their combination treatment for 24 h. After 24 h of total treatment, the cells were incubated at 37°C with 1 mg/mL MTS reagent (Sigma, St. Louis, MO, USA) for 2 h. The formazan crystals were dissolved in isopropanol. Spectrophotometric absorbance of the samples was determined by the ULTRA Multifunctional Microplate Reader (ELx800-BIO-TEK) at 490 nm. MTS assay repeated at least three times.
Mts reagent
Manufactured by Merck Group
Sourced in United States
The MTS reagent is a laboratory product manufactured by Merck Group. It is a colorimetric assay used for the quantitative determination of cell viability and proliferation. The reagent contains a tetrazolium compound that is reduced by metabolically active cells, producing a colored formazan product that can be measured spectrophotometrically.
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Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Lyophilized native chicken type 2 collagen cii, by Merck Group (2 mentions)
Tartrate resistant acid phosphatase staining kit, by Merck Group (2 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Pi rnase, by BD (1 mentions)
Modfit lt software, by BD (1 mentions)
Brdu assay, by Merck Group (1 mentions)
Accuri c6, by BD (1 mentions)
Sb202190, by Merck Group (1 mentions)
Pgc 1α antibody, by Abcam (1 mentions)
Antibodies against, by Cell Signaling Technology (1 mentions)
Compound c, by Merck Group (1 mentions)
Celltiter 96 one solution cell proliferation assay, by Promega (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
A spectrophotometer, by PerkinElmer (1 mentions)
Epoch 2, by Agilent Technologies (1 mentions)
Epoch spectrophotometer, by Agilent Technologies (1 mentions)
Reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
17 protocols using mts reagent
1
Evaluating EGCG and Cisplatin Effects on HeLa Cells
2
Colorimetric Assay for Cell Viability
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A 3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium (MTS) colorimetric assay was used to determine the viability of cells. In brief, cells were seeded in 96-well plates at a concentration of 104 cells per well and incubated overnight to allow cells to attach. After incubation, the cells were treated with lycopene alone (10 µM), cisplatin alone (1 µM), and in combination for 72 h. The cells were incubated at 37 °C in 5% CO2 for 2 h after 72 h of total treatment. For assay a 1 mg/mL MTS reagent (Sigma, St. Louis, MO, USA) was added to each well. The absorbance of the solution was estimated using a microplate reader (BioTek Instruments, Winooski, VT, USA) at 490 nm. The sample readings were calculated by subtracting the mean of background absorbances. Viability was calculated concerning control cells (%). MTS assay was performed at least four times.
3
Colorimetric MTS Assay for Cell Proliferation
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Cell proliferation was measured using the colorimetric MST assay using methanethiosulfonate (MTS) reagent (Sigma Aldrich, St Louis, MO, USA). Briefly, the cells were collected and seeded into 96-well plates in triplicate at 800 cells/well. After 24 h, 20 μL of MTS (5 mg/mL) was added to quantify cell proliferation on seven consecutive days. The cells were incubated with MTS for 3 h and then the optical absorbance of each well was measured at 490 nm using a microplate reader. Cell growth curves were created by plotting the absorbance (ordinate) against time (abscissa).
4
Cell Proliferation and Drug Sensitivity Assay
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Cell proliferation was measured by BrdU assay (Millipore, USA). For cell cycle analysis, after fixation in 70% ethanol, cells were incubated in PI/RNase (BD Biosciences, USA), and analyzed using an Accuri C6 flow cytometer and ModFit LT software (BD Biosciences, USA). For drug sensitivity assay, cells were treated with 0.4 mg/ml 5-FU, 0.004 mg/ml DDP and 0.0004 mg/ml ADM (sigma, USA) for 48 hours. MTS reagent (Sigma, USA) was added to each well following the manufacturer’s instructions to determine drug sensitivity.
5
MTS Assay for Microglial Viability
6
Signaling Pathway Analysis of Myogenesis
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MTS reagent, SB202190, and compound C were purchased from Sigma-Aldrich (St Louis, MO, USA). Antibodies against myosin heavy chain (MHC), Akt, p38 MAPK, and myogenin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against phospho-Akt, phospho-p38 MAPK, and phosphor-AMPK were obtained from Cell Signaling (Beverly, MA, USA). PGC-1α antibody was purchased from Abcam (Cambridge, MA, USA).
7
Cell Proliferation Assay with EPZ005687 and UNC1999
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Viable cell densities were determined by using the CellTiter 96 One Solution Cell Proliferation Assay (Promega, Madison, WI, USA) according to the manufacturer’s instructions. Briefly, cells were plated at a density of 10, 000/well in 96-well plates and incubated for 24–96 h with increasing concentrations of EPZ005687 or UNC1999. The cells were then incubated with MTS reagent (Sigma, M2128) for 3 h, and analyzed with 490 nm absorbance using microplate reader (Bio-Rad, Laboratories, USA). The data analysis was performed by GraphPad Prism (version 5.01, GraphPad Software, USA). The studies were independently conducted in at least three separate times and each experiment had six replicate wells on one plate.
8
Bicalutamide's Effect on Cell Viability
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LetR cells were steroid-depleted for 72 hours and then seeded into a 24-well plate prior to the addition of bicalutamide (Bica) (1 μM) (Sigma Aldrich) or vehicle (dimethyl sulfoxide DMSO; 0.01 %) to regular growth medium. MTS reagent (Sigma Aldrich) was added after 2, 3 and 4 days respectively and the resultant colorimetric outputs analyzed by measuring the absorbance at 490nm using a spectrophotometer (Perkin Elmer).
9
Cell Proliferation Evaluation using MTS Assay
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A density of 800 transfected cells per well were plated and cultured in 96-well plates. Cell proliferation was evaluated for 7 consecutive days using the methanethiosulfonate (MTS) assay. In brief, 20 μL MTS reagent (5 mg/mL, Sigma-Aldrich, Shanghai, China) was added to the plated cells and the optical density (OD) was recorded at 490 nm by a spectrophotometric reader (EPOCH2, Bio-Tek, Winooski, USA) after 3 hrs’ incubation. All experiments were repeated in triplicate.
10
Cell Viability Assay Using Sunitinib
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Cells were seeded onto 96-well plates at a density of 4 × 103 cells per well in media with 10% FBS and allowed to attach for 24 h. DMSO or sunitinib was added at different concentrations. After 24 h of treatment, MTS reagent (Sigma-aldrich St Louis, MO) was added to cells (1:10 ratio) in each well and allowed to incubate at 37°C in a humid atmosphere with 5% CO2 for 2hrs. After that, plate was read at 490 nm using an Epoch spectrophotometer (BioTek Winooski, VT). All experiments were performed in triplicate and repeated a minimum of three times. Percent viability and cytotoxicity as well as standard deviations and IC50 were calculated from the absorbance values using Microsoft Excel.
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