Cell viability was measured using an MTT assay kit (Amresco, USA) as per the manufacturer’s instructions. Briefly, the MTT solution was added to the cultured cells, and following incubation at 37 °C for 4 hours, the supernatants were removed and the formazan crystals were dissolved in 150 μl DMSO. The color was allowed to develop for ten minutes, and the absorbance was measured at 490 nm. The experiment was repeated three times, and the method has been described in our previous study34 (link).
Mtt assay kit
Manufactured by Avantor
Sourced in United States
The MTT assay kit is a colorimetric assay used to measure cell metabolic activity. It provides a quantitative assessment of cell viability and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Multiskan fc, by Thermo Fisher Scientific (2 mentions)
Horseradish peroxidase labeled antibodies, by Santa Cruz Biotechnology (1 mentions)
Cleaved caspase 8, by Abcam (1 mentions)
P eif2α, by Cell Signaling Technology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Cell counting kit 8 cck 8 assay, by Apexbio (1 mentions)
Annexin 5 fluorescein isothiocyanate propidium iodide fitc pi apoptosis detection kit, by Keygen Biotech (1 mentions)
12 protocols using mtt assay kit
1
Cellular Viability Assay with MTT
2
Doxorubicin-Induced Apoptosis Signaling
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B-MD-C1 (wt) was purchased from ATCC, DOX (Adriamycin, ADR) was purchased from Hisun pharmaceuticals, Zhejiang, China and dissolved in sterile saline solution (2 mg/ml) and stored at -20 °C before use. Fetal bovine serum (FBS) and RPMI 1640 medium were purchased from Gibco, USA; Lipofectamine™2000 was purchased from Life Technologies, USA; antibodies against CRT, ERp57, caspase-8 and cleaved-caspase-8 were purchased from Abcam, USA; antibodies against p-PERK, eIF2α and p-eIF2α were purchased from Cell Signaling Technology, USA; β-actin and horseradish peroxidase-labeled antibodies were obtained from Santa Cruz, USA. MTT assay kit was obtained from Amresco, USA.
3
Cell Viability Evaluation by MTT Assay
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Cell viability was determined using an MTT assay kit (Amresco, USA). After treatments cells seeded in 96-well plateswere incubated with MTT solution-containing culture medium for 4 h and formation of the formazan product was measured at 490 nm in a microplate reader.
4
Evaluating PTEC Cytotoxicity with SUA
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Growth-arrested PTECs were seeded onto 96-well plates (0.25×105 cells per well) and exposed to SUA at increasing concentrations (0, 12.5, 25, 50, 100, 150 and 200 µg/ml) for different durations (24, 48 and 72 h). Control cells were treated with blank medium. A commercial MTT assay kit (Amresco, LLC, Solon, OH, USA) was used to measure the cell viability of PTECs. Cells were incubated with MTT solution for 4 h and crystals were dissolved in dimethyl sulfoxide. Cell viability was recorded as a percentage change in the absorbance (570 nm) of treated cells compared with the control cells.
5
Evaluating Cisplatin Cytotoxicity Using MTT Assay
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Cell proliferation and the half maximal inhibitory concentration (IC50) of each cell line to cisplatin were evaluated using an MTT assay kit (Amresco, USA). Cells were seeded into 96‑well plates for 24 hours and treated with or without cisplatin at different concentrations for 48 hours. Then it was incubated with MTT solution-containing culture medium at 37°C for 4 hours and supernatants were then removed. Formazan crystals were dissolved in 150 µL dimethyl sulfoxide (DMSO) with gentle shaking for 10 minutes, and the absorbance value (OD) was measured at 490 nm.
6
MTT Viability Assay for Cell Cultures
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Cell viability was assessed using an MTT assay kit (Amresco, USA) according to the manufacturer’s protocol. The cells were harvested, resuspended in complete medium, and seeded into 96-well culture plates. After culturing for 24, 48, 72, 96, and 120 hr, the MTT solution was added and the cells were incubated at 37°C for another 4 hrs. The supernatant was aspirated, and the formazan crystals were dissolved in 150 μL DMSO. After a 10-mins incubation at 37°C, the absorbance at 490 nm was measured. Each sample was tested in five replicate wells, and the experiment was repeated thrice.
7
Cell Viability Assay Protocol
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The Cell Counting Kit-8 (CCK-8) assay (APExBIO, Houston, TX, USA) or the MTT assay kit (Amresco, Boise, ID, USA) could detect cell viability based on the manufacturer’s instructions using 96-well plates with 2000 cells per well.
8
Promastigote and Amastigote Growth Inhibition
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The promastigote growth inhibition assay was performed as previously described [21 (link)]. Promastigotes were harvested during the exponential growth phase. They were incubated in the presence and absence of several concentrations of LQOFG-2, LQOFG-6, LQOFG-7 (500 to 7.8 µM), and AmB (10 to 0.078 µM) as the positive control. The plate was incubated at 26 °C for 72 h in a biological oxygen demand (B.O.D.) incubator using a Schneider’s medium (Schneider’s Insect Medium 24.5 g/L; L-glutamine 1.8 g/L; glucose 2 g/L and sodium bicarbonate 0.4 g/L; Sigma-Aldrich—St. Louis—USA) supplemented. The growth inhibition was evaluated using an MTT assay kit (Amresco, Solon, OH, USA) according to the manufacturer’s protocol. After 4 h of incubation, 10% sodium dodecyl sulfate was added to dissolve the formazan crystals. The absorbance (540 nm) was measured using a plate reader (Biosystems model ELx800; Curitiba, PR, Brazil). The same procedure was followed to assess inhibition of the axenic amastigote form using these compounds with the following modifications: the treatment time was shortened to 24 h, and the test temperature was raised to 37 °C at a pH of 5.5. Three independent experiments we performed in triplicate.
9
Cytotoxicity and Apoptosis Assay Protocol
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Reagents purchased for the present study were 0.25% trypsin-0.02% EDTA (Jinuo Biomedical Technology Co., Ltd., Hangzhou, China), fetal bovine serum (FBS), Roswell Park Memorial Institute (RPMI) 1640 medium (both from Hangzhou Sijiqing Biological Engineering Co., Ltd., Hangzhou, China), an MTT assay kit (Amresco LLC, Solon, OH, USA), an Annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI)apoptosis detection kit (Nanjing KeyGen Biotech Co., Ltd., Nanjing, China). All other reagents were manufactured domestically and analytically pure.
10
MTT Viability Assay Protocol
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Cell viability assessed using a 3-(4,5-dimethylthiazol-2-yl)-2, 5-diph-enyltetrazolium bromide (MTT) assay kit (AMRESCO). Cells (5×103 cells/well) were seeded into 96-well plates and cultured for 24, 48, 72and 96h. MTT (20μl from a 5mg/ml stock) was added to each well and the plates were incubated at 37ºC for 4h. At the end of incubation, a sodium dodecyl sulfate/isobutanol/HCl solution (100μl/well) was added. The absorbance was measured using a Multiskan™ FC (Thermo Scientific) at 570nm.
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