The effects on the viability of hDPSCs treated with CurLIP, HEMA 3 and HEMA 5 mmol L–1 were evaluated by means of the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliumbromide (MTT) method. 2 × 103 cells/well were placed in a 96-well tissue culture plates and incubated at 37°C for 24, 48 and 72 h. At each time point, MTT solution (20 μl; Promega, Milan, Italy) was added to each well to detect the metabolic activity of the cells. All plates were cultured in the dark for 3 h at 37°C (Diomede et al., 2016a (link)). Supernatants were read at 650 nm wavelength using a microplate reader (Synergy HT, BioTek Instruments, Winooski, VT, United States; Cavalcanti et al., 2015 (link)).
Mtt solution
Manufactured by Promega
Sourced in United States, Italy, China
MTT solution is a colorimetric assay reagent used to measure cell viability and proliferation. The solution contains the tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), which is reduced by metabolically active cells to form insoluble purple formazan crystals. The amount of formazan produced is proportional to the number of viable cells, and can be quantified by spectrophotometric analysis.
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Lab products found in correlation
Penicillin streptomycin, by Lonza (9 mentions)
Fetal bovine serum (fbs), by Merck Group (7 mentions)
L glutamine, by Lonza (5 mentions)
Prism 7, by GraphPad (4 mentions)
Synergy ht, by Agilent Technologies (3 mentions)
Spectramax microplate spectrophotometer, by Molecular Devices (3 mentions)
Microplate reader, by Agilent Technologies (3 mentions)
Rpmi 1640 dmem media l glutamine, by Lonza (2 mentions)
Prism 5, by GraphPad (2 mentions)
Microplate reader, by Bio-Rad (2 mentions)
Fitc annexin 5 apoptosis detection kit with pi, by BioLegend (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
Ccl 247, by American Type Culture Collection (1 mentions)
Imark 168 1130, by Bio-Rad (1 mentions)
Dmem f12, by Merck Group (1 mentions)
Crl 1831, by American Type Culture Collection (1 mentions)
Edu assay kit, by RiboBio (1 mentions)
Mem culture medium, by Thermo Fisher Scientific (1 mentions)
Automatic microplate reader, by Agilent Technologies (1 mentions)
Pancreatin, by Thermo Fisher Scientific (1 mentions)
41 protocols using mtt solution
1
Viability of hDPSCs Treated with CurLIP, HEMA 3, HEMA 5
2
Cell Viability Assay for Compound Screening
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Cells were plated at a density of 5000 cells in triplicates in a 96-well plate. On the next day, treatment of the cells was performed with the indicated compound/s at the specified concentrations in 100 μL media as a final volume. Cell viability was considered after 72 h using MTT solution (Promega, Madison, WI, USA) [28 (link)]. The reagent (20 μLs) was added to each well followed by incubation of the plate for 3 h and fluorescence was subsequently measured (570 nm) using a plate reader, then IC50 values were assessed using the GraphPad prism 7 [29 (link),30 (link)].
3
Cytotoxicity Evaluation of Compounds
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PC-3 and WISH cell lines were obtained from the National Cancer Institute in Cairo, Egypt, cultured in “RPMI-1640/DMEM media l -Glutamine (Lonza Verviers SPRL, Belgium, cat#12-604F). The cells were cultured in 10% fetal bovine serum (FBS, Sigma-Aldrich, MO, USA) and 1% penicillin-streptomycin (Lonza, Belgium)”. All cells were incubated at “37 °C in 5% carbon dioxide atmosphere (NuAire)” following routine tissue culture work. Cells were seeded in triplicate at a density of 5 × 104 cells per well in a 96-well plate, and then treated with the compounds at concentrations of 0.1, 1, 10, and 100 μM the next day. Cell viability was assessed using MTT solution (Promega, USA).60 (link) The plate was incubated for three hours. The absorbance was then measured with an ELISA microplate reader (BIO-RAD, model iMark, Japan). GraphPad Prism 7 was used to determine IC50 values based on survival rates relative to a control group, as previously reported in ref. 61 (link).
4
MTT Assay for Cell Viability
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Two thousand five hundred HDFs were grown per well in FBM containing half of FGM-2 SingleQuots mixed with either an FBM (control group) or an FPF supernatant (FPF group) at 37 °C for up to 4 days. Twenty microliters of MTT solution (Promega, Madison, WI, USA) per 100 μL of medium was added into cell cultures and incubated for 2 h at 37 °C. The absorbance of each well was measured at 490 nm using a plate reader. Data were obtained from at least three different experiments in quintuplicate.
5
Cytotoxicity of Baicalin in Vero Cells
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Cytotoxicity of baicalin against Vero cells was determined using the MTT assay as previously describe30 (link). Briefly, a confluent monolayer of Vero cells in 96-well cell culture microplate were treated with increasing concentrations of baicalin in triplicates for 4 days equivalent to that used in the antiviral assays. After 4 days, MTT solution (Promega, Madison, WI, USA) (15 μl) was added to each well and the microplate was kept at 37°C for 4 h in a humidified atmosphere with 5% CO2. Then, solubilization/stop solution (100 μl) was added to the wells and the absorbance values of the wells were measured at 570 nm using a 96-well plate reader (TECAN, Mannendorf, Switzerland). Dose-response curve was plotted using Graph Pad Prism 5 (Graph Pad Software Inc., San Diego, CA, USA, 2005) and the half maximal cytotoxic concentration (CC50) of baicalin was determined from the plot. Results were represent as the means ± standard error of the mean (SEM) from triplicate assay from three independent experiments.
6
Curcumin Cytotoxicity Assay Protocol
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Cell viability was analyzed by the MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide] assay. Briefly, cells were seeded in 96-well plates at the density of 2×103 cells/well and were cultured with increased concentrations of curcumin for up to 72 h, and then 10 μl of 10 mg/ml MTT solution was added to each well for an additional 4 h according to the manufacturer’s instructions (Promega, Shanghai, China). After centrifugation, 150 μl dimethyl sulfoxide was added to the precipitate and the absorbance of the enzyme was measured at 570 nm using a Microplate Reader (Bio-Rad, Hercules, CA, USA). Cell growth rates (average absorbance of each treat group and non-treated group) were then calculated. All experiments were performed in triplicate and repeated at least three times.
7
MTT Assay for Cell Viability
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Briefly, H1581 or A549 cells were transfected and cultured for 0 h, 24 h, 48 h, or 72 h. Then, 20 μL MTT solution (5 mg/mL; Promega, Madison, WI, USA) was added to cell-culture medium for 4 h. Next, the product was dissolved with 200 μL dimethyl sulfoxide (DMSO). Finally, the optical density (OD) of samples was measured using a microplate reader (Bio-Rad, Richmond, CA, USA) at 490 nm.
8
Paclitaxel Cytotoxicity Assay
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Cells were cultured overnight in 96-well flat bottomed microtiter plates (5 × 103 cells/well) and exposed to 0, 10, 20, 40, 80, 100, 200, or 250 μM paclitaxel. After 0, 24, 48, or 72 h, 20 μl MTT solution (Promega Corporation) was added to each well and cultured at 37°C in 5% CO2 for 4 h. A total of 100 μl formazan solution was added to each well, and the cells were incubated at 37°C in 5% CO2 for 4 h. Absorbance at 570 nm was measured using a spectrometer. GraphPad Prism version 5 (GraphPad Software, Inc.) was used to assess the relative viability of the cancer cells using the following formula: [Control optical density (OD) − experimental OD]/Control OD. Each treatment was performed in quintuplicate.
9
Proliferation Assay of Melanoma Cells
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The proliferation of B16 cells and that of B16/BL6 cells were determined by a 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cells were seeded at 1×103 per well in 96-well plates with the CM and incubated for 72 hr. Subsequently, 100 μl of culture medium and 20 μl of MTT solution (Promega, Tokyo) were added to each well, and the absorbance at 570 nm was analyzed using a microplate reader (Bio-Rad 550; Bio-Rad, Tokyo).
10
Viability Evaluation of hPDLSCs Using MTT Assay
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The viability of the hPDLSCs was determined using the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliumbromide (MTT) method. Two lots of 103 cells/well were placed in a 96-well tissue culture plate and incubated at 37 °C for 24, 48, 72 h and 1 week. At each time point, MTT solution (20 µL) (Promega, Milan, Italy) was added to each well to detect the metabolic activity of the cells [17 (link),18 (link)].
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