The cells were seeded into 96-well plates (1 × 103 cells in each well) and treated as indicated for 96 h. Cell viability was assessed using the CellTilter 96® Aqueous One Solution Cell Proliferation Assay (Promega; Madison, WI, USA), according to the manufacturer’s instructions.
Celltilter 96 aqueous one solution cell proliferation assay
Manufactured by Promega
Sourced in United States
The CellTiter 96® Aqueous One Solution Cell Proliferation Assay is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. The assay involves the use of a tetrazolium compound (MTS) that is bioreduced by cells into a colored formazan product that is soluble in tissue culture medium. The absorbance of the formazan product is directly proportional to the number of living cells in culture.
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Lab products found in correlation
Microplate reader, by Agilent Technologies (2 mentions)
Synergy mx, by Agilent Technologies (1 mentions)
Spectramax i3x plate reader, by Molecular Devices (1 mentions)
I3x sc acad, by Molecular Devices (1 mentions)
1640 glutamax 1, by Thermo Fisher Scientific (1 mentions)
Hyclone, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Multiskan go microplate spectrophotometer, by Thermo Fisher Scientific (1 mentions)
9 protocols using celltilter 96 aqueous one solution cell proliferation assay
1
Cell Viability Assay in 96-well Plates
2
Evaluating Cell Viability with Cy3-Labeled Probes
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AGS cells were seeded in 96-well plates with a final volume of 200 μL of complete medium per well and grown to 50% and 100% confluence. Upon reaching the desired confluence, AGS cells were incubated with 200 nM of Cy3_ HP_ LNA/2OMe _PS diluted in hybridization buffer [0.5 M urea and 900 mM NaCl; 1% (v/v) or 5% (v/v)] or in complete medium (untreated control) for 24 hours. At the end of the incubation period, the cell viability was evaluated using CellTilter 96 Aqueous One Solution Cell Proliferation Assay (Promega Corporation, Madison, WI), according to the manufacturer’s instructions. Absorbance at 490 and 630 nm was measured using a microplate reader (Biotek Instruments Inc. Synergy MX, USA). Background absorbance values were subtracted from the absorbance values generated with MTS assay. The values from treated cells were compared with the values generated from untreated control cells and reported as percent viability. All experiments were performed in triplicate.
3
Nrf2 Knockdown Impacts Cell Viability
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The cells were seeded at a density of 1-2 × 104 cells/well in a 96-well plate and transfected with siNrf2 for 48 hours. Cell viability was assessed by Cell Tilter 96 Aqueous One Solution Cell Proliferation Assay (Promega Corporation, Madison, WI), according to the manufacturer's instructions. Untreated cells served as a negative control. The experiments were performed in triplicate.
4
Cell Proliferation MTS Assay
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2- 4 × 103 cells were seeded in 96-well plate in 100 µl of medium per well. After 72 h, MTS assays were performed according to the protocols of Cell Tilter 96 aqueous One Solution Cell Proliferation assay (Promega). The absorbance at 490 nm was measured using a spectraMax i3X plate reader (Molecular Devices, I3X-SC-ACAD).
5
Cell Viability Assay for AGS Cells
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The human gastric epithelial cell line AGS (ATCC® CRL-1739) was cultured in RPMI medium 1640 Glutamax I (Gibco, Invitrogen, Grand Island, NY, USA) supplemented with 10% (v/v) fetal bovine serum (FBS, HyClone, Thermo Fisher Scientific, Inc, UK) and 1% (v/v) Penicillin/Streptomycin (Gibco) at 37°C in a humidified 5% CO2 atmosphere. Cells were seeded at 1.6 x 105 cells/well in a 96-well plate. After overnight, media were changed and cells were incubated with predetermined concentrations (0.4 μM to 2 μM) of Cy3_HP_ LNA/2OMe _PS probe diluted in vehicle (0.5 M urea and 900 mM NaCl) or only with the vehicle, for 24 h. Cell viability was assessed by CellTilter 96® Aqueous One Solution Cell Proliferation Assay (Promega Corporation, Madison, WI), according to the manufacturer’s instructions. Untreated cells served as a negative control. The experiments were performed in triplicate.
6
Proliferation Assay for 3D Cell Spheroids
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A total of 1.5 × 104 HCT116 cells were seeded in 96-well low attachment plates in SCM, and then newly formed spheroids were treated with BK and NT analogues in duplicate. The entire experiment was repeated three times. Non-treated cells were used as a negative control. After 3 days, cell proliferation was assessed by a colorimetric CellTilter 96® Aqueous One Solution Cell Proliferation Assay (Promega Corporation, Madison, WI, USA), according to the manufacturer’s instructions. Briefly, 20 μL of CellTiter 96® AqueousOne Solution Reagent was added into each well of the 96-well assay plate containing the samples in 200 μL of culture medium. After 4 h of incubation, the absorbance at 490 nm was recorded using a microplate reader (Epoch, BioTek Instruments, Winooski, VT, USA).
The same test was used in the preliminary experiments with adherent HT29 cells.
The same test was used in the preliminary experiments with adherent HT29 cells.
7
Cell Viability Assay Protocol
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Cell viability was assessed by using the CellTilter 96 Aqueous One Solution Cell Proliferation Assay (Promega Corporation, Madison, WI, USA), according to the manufacturer’s instructions. Cells were seeded at a density of 1 × 104 cells/well into 96-well plates. After their subjection to different treatments, the cells were incubated with 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H–tetrazolium, inner salt (MTS) solution at a final concentration of 0.4 mg/ml for 4 h at 37 °C. One Solution Reagent was then added directly to the culture wells, and the plates were incubated for 4 h, following which the absorbance at 490 nm was recorded with a 96-well plate reader [17 (link), 18 (link)]. The absorbance was measured at 490 nm with a Multiskan GO microplate spectrophotometer (Thermo Fisher Scientific).
8
Spheroid Cell Proliferation Assay
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HCT116 and HT29 cells (1.5×104/ml were seeded in 96-well low attachment plates in SCM, then newly formed spheroids were treated with T3 (0.5, 1 or 2 nM) in 3 replicates for each option. The whole experiment was repeated twice. Non-treated cells were negative control. After 3 days, cell proliferation was assessed by colorimetric Cell Tilter 96® Aqueous One Solution Cell Proliferation Assay (Promega Corporation), according to the manufacturer's instructions. Briefly, 20 µl of Cell Titer 96® Aqueous One Solution Reagent were added into each well of the 96-well assay plate containing the samples in 200 µl of culture medium. After 4 h of incubation at 36°C the absorbance at 490 nm was recorded using a microplate reader (BioTek Instruments, Inc.).
9
In vitro DC-Effector Cell Proliferation
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The in vitro modified DCs and specific effector cells (mentioned above) were put in triplicate on 96-well plate in different proportions, ranging from 0.0625 to 2 (DC/effector cell number) and incubated for 24 h. After that time, cell proliferation was assessed by colorimetric CellTilter 96® Aqueous One Solution Cell Proliferation Assay (Promega Corporation), according to the manufacturer’s instructions. Briefly, 20 μl of CellTiter 96®AqueousOne Solution Reagent were added into each well of the 96-well assay plate containing the samples in 200 μl of culture medium. After 4 h of incubation the absorbance at 490 nm was recorded using a microplate reader (BioTek, Winooski, USA).
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