A2058 and HT144 cells were treated with vehicle control or I3A at indicated concentrations for 48 h in 96-well plates. Cell viability was assessed by using the MTT cell viability assay kit (Thermo Fisher, USA) as per the manufacturer’s instructions. After 48 h of treatment, 20 μl of MTT reagent (2 mg/ml) was added to wells containing cells in the plate. Plates were then incubated at 37°C for 2 h. Then, media from wells containing MTT reagent were completely aspirated and wells were dried. Then, 100 μl of DMSO was added to each well to dissolve the formazan crystals formed, and absorbance was recorded at 570 nm in a microplate reader. The cell viability ratio was calculated and values are presented as percentage of control. For clonogenic assay, A2058 and HT144 cells were treated with vehicle control or with I3A at 1 and 5 μM concentrations for 24 h. Then, media were removed and plates were incubated with methanol for 30 min. Cells were then stained with crystal violet for 60 min followed by washing with phosphate-buffered saline (PBS). Then, cell counting was performed under a light microscope (40×). The experiment was performed in triplicate with a minimum of 3 wells per sample in each experiment.
Mtt cell viability assay kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The MTT Cell Viability Assay Kit is a colorimetric assay used to measure the metabolic activity of cells. It utilizes the yellow tetrazolium salt MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), which is reduced by metabolically active cells to form purple formazan crystals. The resulting color change is proportional to the number of viable cells and can be quantified using a spectrophotometer.
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Lab products found in correlation
Microplate absorbance reader, by Bio-Rad (3 mentions)
Tgf β1, by R&D Systems (1 mentions)
Spectramax id3, by Molecular Devices (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Reverse transcriptase kit, by Thermo Fisher Scientific (1 mentions)
Ab224736, by Abcam (1 mentions)
Ab8245, by Abcam (1 mentions)
9 protocols using mtt cell viability assay kit
1
Cell Viability and Clonogenic Assay
2
Evaluating PTD-BMP-7 Effects on HPMC Viability
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3‐(4,5 dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) cell viability assay kit (Thermo Fisher Scientific) was performed to determine the PTD‐BMP‐7 dose used in the experiments.20 Subconfluent HPMCs were FBS‐restricted for 24 hours after which the media was replaced into 1% FBS media for the control group and the same media with TGF‐β1 (2 ng/mL) (R&D Systems) for the TGF‐β1 group. Both groups were treated with PTD‐BMP‐7 (100 ng/mL). Preliminary experiments determined the doses of TGF‐β1 and PTD‐BMP‐7 in this study. Cells were harvested 72 hours after treatment with each stimulus.
3
Ginsenoside-Rg3 Antiproliferative Assay
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We measured cell proliferation by MTT assay as described by Kumar et al. [21 (link)]. The transfected SMMC-7721 and SK-Hep-1 cells were placed into 24-well plates and incubated at 37°C with 5% CO2 for 48 h. Cells were treated with 1, 2, 4, 8, and 16 μg/ml of ginsenoside-Rg3. Subsequently, MTT Cell Viability Assay Kit (Thermo Fisher Scientific, USA) was performed according to the manufacturer's protocol. Moreover, we detected OD value under 570 nm wavelength using a microplate reader at the 0, 12, 24, 48 and 72 h time points.
Then, the lowest cell viability was in the group treated with 8 μg/ml ginsenoside-Rg3; therefore, 8 μg/ml ginsenoside-Rg3 was selected for the following experiments.
Then, the lowest cell viability was in the group treated with 8 μg/ml ginsenoside-Rg3; therefore, 8 μg/ml ginsenoside-Rg3 was selected for the following experiments.
4
HUVEC Viability Assessment via MTT Assay
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The viability of the HUVECs was determined by Methyl tetrazolium (MTT) cell viability Assay Kit (V13154, Thermo Fisher, Massachusetts, USA). The cells were collected and transferred to 96-well plates after adjusting cell concentration (5 × 10 3 cells/well). After the cells in each group were appropriately treated, an appropriate amount (10 μL) 12-mM MTT stock solution was added to each well according to the operating instructions, and then the cells were incubated at 37°C for 4 h. Next, 100 μL SDS-HCl solution was added to each well and the cells were incubated for 4 h. Finally, the absorbance of each sample well at 570 nm was detected with a microplate reader (SpectraMax iD3, Molecular Devices, Calif, USA).
5
Measuring Cell Proliferation via MTT Assay
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Cell proliferation was evaluated using MTT Cell Viability Assay Kit (Invitrogen; Carlsbad, CA, USA) following the manufacturer’s instructions. Briefly, approximately 1 × 104 BcaCD885 and Tca8113 cells were seeded into 96-well plates and treated with different concentrations of flavopereirine for 24, 48 and 72 h. Subsequently, 10 μL of MTT stock solution was added and incubated at 37°C for another 4 h. Then, 100 μL of dissolution reagent was added and incubated for 4 h. Finally, the absorbance at 490 nm was detected under a microplate reader (TECAN-infinite M200pro, Mannedorf, Switzerland).
6
MTT Cell Viability Assay Protocol
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Cell viability was evaluated by using the MTT Cell Viability Assay Kit (Invitrogen; Carlsbad, CA, USA) according to the manufacturer’s instructions. Briefly, after transfection and OGD/R treatment, 10 μl of MTT stock solution was added and incubated for 4 h at 37℃. Then, 100 μl of dissolution reagent was added for another 4 h. Finally, the absorbance at 490 nm was measured with a microplate absorbance reader (Bio-Rad, Sunnyvale, CA, USA).
7
MTT Cell Viability Assay Protocol
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Cell viability was evaluated using an MTT Cell Viability Assay Kit (Invitrogen; Thermo Fisher Scientific, Inc.), according to the standard protocols. In brief, transfected or treated HT22 cells cultured in 96-well plates were added to 10 μL of MTT stock solution for 4 h at 37 °C. Then, 100 μL of dissolution reagent was added to each well and incubated for another 4 h. The absorbance was detected using an optical density measurement at 570 nm under a microplate absorbance reader (Bio-Rad, Sunnyvale, CA, USA). Cell viability of the control group was regarded as 100% and cell viabilities in other groups were calculated as percentages of the control.
8
Investigating miR-488-3p and VPS4B Roles
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The miR-488-3p mimic and its negative control (NC), miR-488-3p inhibitor and its negative control (NC), and lentivirus-mediated VPS4B shRNA (sh-VPS4B) and its negative control (sh-NC) were obtained from Ribobio (Guangzhou, China). Trizol reagent, the Reverse Transcriptase kit, and MTT Cell Viability Assay Kit were bought from Invitrogen (Carlsbad, CA, USA). The primary antibody including anti-VPS4B (ab224736, Abcam) and anti-GAPDH (ab8245, Abcam) were applied from Abcam. Other reagents and buffers used in this research were all obtained from Sigma-Aldrich.
9
MTT Cell Viability Assay for Assessing Cellular Responses
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MTT Cell Viability Assay Kit (Invitrogen; Carlsbad, CA, USA) was applied to measure the cell viability following the manufacturer’s protocol. Briefly, after being planted into 96-well plates, cells were cultured overnight. After transfection and treatment, the medium was refreshed, and we added MTT stock solution (10 μL) into each well at 37°C for 4 hours. After that, dissolution reagent (100 μL) was added to each well and treated at 37°C for 4 hours. An optical density measurement at 570 nm using a microplate absorbance reader (Bio-Rad, Sunnyvale, CA, USA) was utilized to analyze the absorbance.
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