Cells were grown to 50–60% confluency overnight in 96-well plates. Media was aspirated, and the cells were incubated with fresh media containing either CAHA-sSWCNTs or sSWCNTs or CAHA at various concentrations along with control cells for 48 h. After treatment the cells were washed two times in PBS, and cells were incubated for an additional 24 h in fresh media. MTT was assessed using the CellTiter 96 AQ One Solution cell proliferation assay kit (Promega, MI, USA) and measured optically at 570 nm.
Celltiter 96 aq one solution cell proliferation assay kit
Manufactured by Promega
Sourced in United States
The CellTiter 96 AQ One Solution Cell Proliferation Assay Kit is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. The kit contains a tetrazolium compound that is bio-reduced by cells into a colored formazan product, which can be quantified by absorbance measurements.
Automatically generated - may contain errors
Lab products found in correlation
Inverted microscope, by Olympus (1 mentions)
Cellquest software, by BD (1 mentions)
Elite esp flow cytometry, by BD (1 mentions)
Multilabel counter, by PerkinElmer (1 mentions)
Nnc 55 0396, by Merck Group (1 mentions)
Mibefradil, by Merck Group (1 mentions)
Facscan flow cytometer, by BD (1 mentions)
Verity winlist software, by Verity Software House (1 mentions)
7 protocols using celltiter 96 aq one solution cell proliferation assay kit
1
Cytotoxicity of CAHA-sSWCNTs Evaluated
2
MTS Assay for Cell Proliferation
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Cells were seeded into 96‐well plates. 3‐(4,5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium, inner salt (MTS) assays were performed using the CellTiter 96 AQ One Solution Cell Proliferation Assay Kit (Promega) according to the manufacturer's instructions. The absorbance was measured at 490 nm.
3
Evaluating Cell Proliferation with MTS and EdU
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Cell proliferation was evaluated by MTS and EdU assays. MTS assay was performed using a CellTiter 96 AQ One Solution Cell Proliferation Assay kit (MTS, Promega, Madison, USA) according to the manufacturer’s instructions. Cells were cultured in 96-well plates at 1 × 104 cells per well. Transfected and untransfected cells were treated with 30 mg/L DADS for 24 h or left untreated, and the absorbance was recorded at 490 nm using an ELISA plate reader. Each assay was replicated 5 times.
EdU assay was conducted with an EdU kit (RiboBio, Guangzhou, China). The untreated and treated cells were incubated with the diluted EdU medium for 2 h, and then washed in PBS and subjected to DAPI staining. Finally, the images were taken and analyzed using an inverted microscope (Olympus, Tokyo, Japan). All the assays were repeated three times.
For cell cycle analysis, cells were fixed in ice-cold 70% ethanol and stained with propidium iodide (PI). The cell cycle profiles were assayed using Elite ESP flow cytometry at 488 nm and the data were analyzed using CELL Quest software (BD Biosciences, San Jose, CA, United States).
EdU assay was conducted with an EdU kit (RiboBio, Guangzhou, China). The untreated and treated cells were incubated with the diluted EdU medium for 2 h, and then washed in PBS and subjected to DAPI staining. Finally, the images were taken and analyzed using an inverted microscope (Olympus, Tokyo, Japan). All the assays were repeated three times.
For cell cycle analysis, cells were fixed in ice-cold 70% ethanol and stained with propidium iodide (PI). The cell cycle profiles were assayed using Elite ESP flow cytometry at 488 nm and the data were analyzed using CELL Quest software (BD Biosciences, San Jose, CA, United States).
4
Assessing Cell Viability and Proliferation
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The number of viable cells was counted by trypan blue exclusion using a hemocytometer. Cells (1 × 104) were seeded in 96-well plates and transfected with non-silencing siRNA and siPALMD-1. In the following day, cells were treated with 0.2 μg/ml ADR. MTS assays were performed using the CellTiter 96 AQ One Solution Cell Proliferation Assay Kit (Promega) following the manufacture's protocol. The absorbance was measured at 490 nm with use of a multilabel counter (PerkinElmer).
5
Cell Viability Assay with Mibefradil and NNC-55-0396
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To determine cell survival and proliferation, cell growth was quantified using the CellTiter 96 AQ One Solution Cell Proliferation Assay Kit (Promega, Madison, WI, USA). Cells were plated in 96-well culture plates at a density of 1–2 × 104 cells/well in 100 μL of cell culture media. Cells were treated with different concentrations of mibefradil or NNC-55-0396 (Sigma-Aldrich, St. Louis, MO, USA). After drug exposure, 20 μL of CellTiter 96 AQ One Solution Reagent was added to each well and allowed to incubate for 2 h at 37 °C. The quantity of formazan product formed, which is directly proportional to the number of viable cells, was measured on a Multi-Mode Microplate Reader (MD SpectraMax M3, CA, USA) at 490 nm wavelength using a reference filter at 650 nm wavelength. Viability assays were performed at least three times in independent experiments, using triplicate measurements in each.
6
Cell Proliferation on Zn-Ti Surfaces
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Cells were seeded at a density of 1500 cells/cm2 on control and Zn-Ti samples and were cultured at 37 °C in a humidified atmosphere with 5% CO2.
Proliferation of seeded cells cultured for 1, 3, 5, and 7 days was detected using the CellTiter 96 AQ One Solution Cell Proliferation Assay Kit (Promega, USA) in accordance with the manufacturer’s instructions. The absorbance of each sample was observed at 562 nm using a plate reader (Varioskan Frash 2.4; Thermo science, USA).
Proliferation of seeded cells cultured for 1, 3, 5, and 7 days was detected using the CellTiter 96 AQ One Solution Cell Proliferation Assay Kit (Promega, USA) in accordance with the manufacturer’s instructions. The absorbance of each sample was observed at 562 nm using a plate reader (Varioskan Frash 2.4; Thermo science, USA).
7
Cell Proliferation and Cell Cycle Analysis
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Cell proliferation assays were performed using a CellTiter 96 AQ One Solution Cell Proliferation Assay kit (MTS, Promega, Madison, USA) according to the manufacturer's instructions. Cells were seeded in 96-well plates at 1 × 104 cells per well. Transfected and untransfected cells were treated with 30 mg/L DADS for the indicated times or left untreated, and the absorbance was recorded at 490 nm using an ELISA plate reader. Each assay was replicated 5 times.
Cell cycle analyses were conducted as follows. Briefly, transfected and untransfected cells were treated with 30 mg/L DADS for 24 h or left untreated. Cells were harvested and resuspended in ice-cold 75% ethanol and then fixed for 24 h at 4°C. For subsequent flow cytometry analysis, fixed cells were resuspended in 1 mL of PI (propidium iodide) staining reagent for 30 min. The data were collected using a FACScan flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA) and analyzed with Verity Winlist Software (Verity Software House, Topsham, ME, USA).
Cell cycle analyses were conducted as follows. Briefly, transfected and untransfected cells were treated with 30 mg/L DADS for 24 h or left untreated. Cells were harvested and resuspended in ice-cold 75% ethanol and then fixed for 24 h at 4°C. For subsequent flow cytometry analysis, fixed cells were resuspended in 1 mL of PI (propidium iodide) staining reagent for 30 min. The data were collected using a FACScan flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA) and analyzed with Verity Winlist Software (Verity Software House, Topsham, ME, USA).
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