The CAECs at passage two or three were dispersed into a cell suspension and seeded in a 96-well plate at a density of 5 × 104 cells/well (6 parallel wells for each group). Upon attaining 80% cell confluence, the transfected cells were collected and incubated in 20 μL of MTT solution (Sigma-Aldrich Chemical Company, St Louis, MO, USA) at 37° C for 4 h. After removal of the MTT solution, the cells in each well were treated with 150 μL of dimethyl sulfoxide (Sigma-Aldrich Chemical Company, St Louis, MO, USA) in a shaker for 10 min. The OD of each well was measured at an excitation wavelength of 490 nm using a microplate reader. The average OD values were determined from the mean of three independent measurements. The cell viability was calculated according to the following equation: (OD the experimental group - OD the control group)/OD the control group.
Mtt solution
Manufactured by Merck Group
Sourced in United States, Germany, Sao Tome and Principe, Italy, China, United Kingdom, France, Spain, Canada, Macao, Australia
MTT solution is a biochemical assay used to measure cell viability and proliferation. It is a colorimetric assay that relies on the conversion of the yellow tetrazolium salt MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) into purple formazan crystals by the mitochondrial enzymes of living cells. The amount of formazan produced is directly proportional to the number of viable cells, which can be quantified using a spectrophotometer.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (391 mentions)
Microplate reader, by Bio-Rad (115 mentions)
Microplate reader, by Agilent Technologies (57 mentions)
Microplate reader, by Thermo Fisher Scientific (54 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (32 mentions)
Microplate reader, by Molecular Devices (29 mentions)
Microplate reader, by Tecan (19 mentions)
Elx800, by Agilent Technologies (18 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (18 mentions)
96 well plate, by Corning (16 mentions)
Multiskan go, by Thermo Fisher Scientific (14 mentions)
Multiskan fc, by Thermo Fisher Scientific (14 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (13 mentions)
96 well plate, by Thermo Fisher Scientific (12 mentions)
Mtt assay, by Merck Group (12 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (11 mentions)
Spectramax plus 384, by Molecular Devices (9 mentions)
Microplate reader, by BMG Labtech (9 mentions)
Versamax microplate reader, by Molecular Devices (8 mentions)
Synergy ht, by Agilent Technologies (8 mentions)
1 604 protocols using mtt solution
1
MTT Cell Viability Assay Protocol
2
Cell Proliferation Measurement via MTT Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MTT assay was applied to measure cell proliferation. Cells were collected at 24 h posttransfection and seeded into 96-well plates with a density of 3,000 cells per well. Then the transfected cells were cultured at 37°C with 5% CO2 for 0, 24, 48, or 72 h. A total of 10 μl of MTT solution (5 mg/ml; Sigma-Aldrich, St. Louis, MO, USA) was added into each well and incubated for an additional 4 h at 37°C. Then the supernatant containing MTT solution was removed, and 150 μl of dimethyl sulfoxide (Sigma) was added into each well. After incubation at 37°C for 20 min, optical density (OD) was determined at a wavelength of 490 nm using a SpectraMax M5 microplate reader (Molecular Devices, LLC, Sunnyvale, CA, USA). Each assay was independently conducted in triplicate.
3
EGCG Cytotoxicity Evaluation in TWNT4 Cells
4
Cell Proliferation Measurement by MTT Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Growth profiles of cells cultured between days 1 to 8 from the two groups were measured by an MTT assay. Briefly, cells at passage 3 were seeded at a density of 1 000 cells per well in 96-well plates containing 10% α-MEM. At each time point, 20 μL of MTT solution (1 mg·mL−1; Sigma-Aldrich) was added to each well; thereafter, all the plates were returned to standard tissue incubator conditions for an additional 4 h. After the removal of the MTT solution, 150 μL dimethyl sulfoxide (DMSO; Sigma-Aldrich) was added to each well and incubated while shaking in the dark for 15 min. The cell proliferation profiles were determined from the measurements obtained at a wavelength of 490 nm with a microplate reader (Shanghai Precision Instrument Co. Ltd., Shanghai, China). Experiments for all samples were repeated in triplicate.
5
MTT Assay to Assess Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MTT assay was performed to monitor cell growth and cell viability at different
time. After H1299 and A549 cells were cultured and transfected with siNC,
siTMPO-AS1, siTMPO-AS1plus miR-143-3p inhibitor and siTMPO-AS1 plus inhibitor
control respectively, 10 µL of MTT solution (5 mg/ml, SIGMA, Saint Louis, US)
was added to each well at 24 h, 48 h and 72 h after transfection. Another set of
experiments were performed by transfecting miR-143-3p inhibitor, inhibitor
control, siNC plus inhibitor or siCDK1 plus inhibitor into LC cells, but MTT
solution was added at 48 h after transfection only. Then cells were incubated at
37°C with 5% CO2 for another 4 h. Next, MTT solution was removed
followed by adding 150 µL of dimethyl sulphoxide (DMSO, SIGMA, Saint Louis, US)
to each well. Optical density (OD) was recorded at a wavelength of 570 nm in a
microplate reader (Bio-Rad, California, US).
time. After H1299 and A549 cells were cultured and transfected with siNC,
siTMPO-AS1, siTMPO-AS1plus miR-143-3p inhibitor and siTMPO-AS1 plus inhibitor
control respectively, 10 µL of MTT solution (5 mg/ml, SIGMA, Saint Louis, US)
was added to each well at 24 h, 48 h and 72 h after transfection. Another set of
experiments were performed by transfecting miR-143-3p inhibitor, inhibitor
control, siNC plus inhibitor or siCDK1 plus inhibitor into LC cells, but MTT
solution was added at 48 h after transfection only. Then cells were incubated at
37°C with 5% CO2 for another 4 h. Next, MTT solution was removed
followed by adding 150 µL of dimethyl sulphoxide (DMSO, SIGMA, Saint Louis, US)
to each well. Optical density (OD) was recorded at a wavelength of 570 nm in a
microplate reader (Bio-Rad, California, US).
6
MTT Viability Assay for OSCC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After transfection, OSCC cells were harvested, seeded into 96-well culture plates at a density of 2000 cells in 200 uL/well, and incubated at 37°C. At different time points (24, 48, 72, or 96 h), 100 uL of MTT solution (0.5 mg/mL; Sigma, USA) was added to each well, and the plates were incubated for another 4 h. Then, the MTT solution was removed and 150 uL dimethyl sulfoxide (DMSO) was added to each well to stop the reaction. The plates were gently shaken on a swing bed for 10 min, and spectrometric absorbance at 490 nm was measured using a microplate reader. This experiment was run in triplicate for each sample.
7
Evaluating A375 Cell Viability via MTT Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Firstly, transfected A375 cells (4×103 cells/well) were prepared in a 96-well plate. A375 cells were then incubated in fresh medium for 24, 48, 72 or 96 h, respectively. After that, 10 μL of MTT solution (Sigma–Aldrich, St. Louis, MO, USA) was added there, and the cells were further cultured for 4 h. Next, the MTT solution was aspirated and the Formazan solution was added to fully dissolve the crystals. The absorbance of each well was measured with a microscope (Olympus Corp., Tokyo, Japan) at 490 nm.
8
MTT Assay for Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh NSC medium 900 μL and MTT solution 100 μL (Sigma) were added into each well of a 96-well plate, and the plate was incubated at 37°C for 3 h. After incubation, the MTT solution was replaced with DMSO (Sigma, United States) and incubated on an orbital shaker for 15 min. The absorbance of the plate was read at OD = 540 nm.
9
Evaluating miR-610's Role in Glioma Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MTT assay was performed to investigate whether miR-610 had a role in glioma cell proliferation. Post-transfection with miR-610 or NC mimics and inhibitors, cells were collected and seeded in 96-well plates at 3,000 cells/well. MTT assay was performed every 24 h for 6 days. Briefly, 20 µl MTT solution (Sigma-Aldrich, St. Louis, MO, USA) was added to each well and incubated for 4 h at 37˚C. The culture medium containing MTT solution was then removed, and 200 µl dimethyl sulfoxide was added to each well. The absorbance was measured at 490 nm using an enzyme-linked immunosorbent assay reader (Bio-Rad Laboratories, Inc., Hercules, CA, USA). All experiments were analyzed in triplicate.
10
Evaluating Cytotoxicity of Zinc Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cytotoxicity of all prepared compounds (LF, Zn-NPs, and LF-Zn-NPs) was tested in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium reduction test (MTT assay) on normal Vero cells and PBMCs cells. 96-well sterile plates were seeded with normal cells (1 × 104 cells/well) and allowed to settle overnight. Serial concentrations (0.0, 125, 250, 500, 1000 and 2000 µg/mL) of the prepared compounds were added to the cells. MTT solution (0.5 mg/mL; Sigma, USA) was added to each well after 48 h of incubation, and the plates were then incubated at 37 °C for about 5 h. Following the removal of the MTT solution, 200 µl of dimethyl sulfoxide (DMSO) was added to each well82 (link). A microplate reader was then used to detect the absorbance at 570 nm (BMG LabTech, Germany). The 100% safe concentration of the preparations (EC100) and half maximum inhibitory concentration (IC50) values were calculated using the Graphpad InStat program 7. Furthermore, the effect of the prepared Zn-NPs and LF-Zn-NPs at concentrations of 250, 500 and 1000 µg/mL on the morphological changes of Vero cells were investigated by phase contrast microscope (Olympus, Japan) in comparison with LF.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!