Vybrant mtt assay kit

The inhibition of mammalian cell viability by the produced EPS was examined by measuring the cell proliferation rate of tumor cells with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay described by Mosmann [72 (link)] where the cancer cells (MDA-MB 231 human breast cancer cell line) were seeded in a 24-well plate at a density of 3 × 10⁴ cells/well. After 24 h, fresh DMEM was added to the cells with increasing concentrations of MOE6-EPS (0, 2, 4, 8 and 16 mg/mL) at 37 °C and 5% CO₂ in air. The growth of the cells was monitored for 96 h and then the cell viability was assessed with the Vybrant MTT assay kit (Thermofisher, Waltham, MA, USA) according to the manufacturer’s suggested protocol. Absorbance (570 nm) of the formazan produced from the reduction of the tetrazolium dye was measured on an Enspire 2300 Multilabel Reader (Perkin Elmer, Waltham, MA, USA). The MTT assay was performed three times with three replicates conducted for each experiment.

Cytotoxicity was determined by using Vybrant® MTT assay kit (Thermo Scientific). Similar to the previous report [20 (link)], RAW 264.7 cells (1 × 10⁶ cells) were treated for 16 h with SKBHT dissolved in PBS, and then metabolically active cells were measured by a plate reader (BioTeK, VT, USA) as instructed by the manufacturer's protocol (Thermo Scientific). Live cells were calculated in a percentage against untreated cells. Each assay was conducted in triplicate samples, and measurement was repeated three times.

Cytotoxicity was determined using a Vybrant® MTT assay kit (Thermo Fisher Scientific, Waltham, MA, USA). Cell culture plates of A549 and SNU2292 were treated with various amounts of eGSM dissolved in PBS for 16 h and measured by a plate reader (BioTeK, VT, USA), as instructed by and the manufacturer. The percentage of live cells was calculated over untreated cells. The assay was conducted in triplicate and repeated three times.

Possible toxicity on the cell, which could be elicited by eCS, was determined by an MTT assay (vybrant MTT assay kit, Thermo Fisher Scientific). Live cells were calculated as described previously [20 (link)]. Each experiment was set in triplicate and performed three times independently.

We used: 2NP, 3NP, and 4NP (99% purity grade), DMSO (Molecular Biology Grade), and Dulbecco’s phosphate-buffered saline (PBS) from Sigma Aldrich (Merck), Poland; high-purity chemicals for gas chromatography/mass spectrometry (GC-MS) from Aldrich Chemical Co. (Milwaukee), USA, used without further purification; solvents for GC-MS (GC² quality) analysis from Burdick and Jackson (Muskegon, MI), USA; and Milli-Q water (18.2 MΩ) from a Millipore Advantage system (Merck), Poland.
We used: Pierce™ lactate dehydrogenase (LDH) cytotoxicity assay kit, Vybrant MTT assay kit (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, calcein-AM, propidium iodide (PI), carboxy-dihydro dichlorofluorescein diacetate (carboxy-H₂DCFDA), MitoSOX™ red, Hoechst 33342 solution, tetramethylrhodamine methyl ester (TMRM), DAPI (4′,6-diamidino-2-phenylindole), and live-cell imaging solution from Invitrogen (ThermoFisher Scientific, USA); HEPES (Bioultra for Molecular Biology Grade) from Merck;, USA Annexin V-FITC kit from Miltenyi Biotec, Spain; and Trypan blue solution and Triton X-100 solutions from Merck, Poland.