To investigate and compare the effect of 24- and 48-hour acidic and neutral LCFSs derived from three Lactobacillus probiotic bacteria (in concentrations from 0% (control) to 50% (v/v)) on the survival and proliferation of PDLSCs, the MTT colorimetric test was used (13 (link)). On the first day of the study, a 96-well plate (SPL Life Sciences, Korea) was seeded with 3,500 cells/well in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, UK) and incubated in a humidified incubator for 24 hours at a temperature of 37°C and 5% CO2. On the second day of the study, cell treatment was performed with different concentrations of LCFS. The control group was the complete cell culture medium alone. Two series of identical plates were treated to check the acute (24 hours) and chronic (72 hours) cytotoxicity of LCFSs on PDLSCs. To perform the MTT assay, the plates corresponding to the designated testing time were removed from the incubator. After washing with phosphate-buffered saline buffer, a cell culture medium containing 10% MTT dye (Sigma-Aldrich, Germany) was introduced in each well. After 2 hours of incubation in a humidified incubator, the MTT dye was replaced with dimethyl sulfoxide (Sigma-Aldrich, Germany). The optical absorption of the resulting color was read at 570 and 620 nm using the plate reader (Anthus 2020, Austria).
Mtt dye
Manufactured by Merck Group
Sourced in United States, Germany, Macao, India, China
The MTT dye is a colorimetric assay used to measure cell metabolic activity. It is a tetrazolium-based compound that is reduced by metabolically active cells, producing a colored formazan product that can be quantified spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (59 mentions)
Microplate reader, by Bio-Rad (5 mentions)
Doxorubicin, by Merck Group (4 mentions)
Synergy h1 hybrid reader, by Agilent Technologies (3 mentions)
Dimethyl sulphoxide, by Merck Group (3 mentions)
L glutamine, by Merck Group (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Victor x3, by PerkinElmer (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions)
Thermo labsystems multiskan rc microplate reader, by Thermo Fisher Scientific (2 mentions)
Calcusyn software, by Biosoft (2 mentions)
Microplate spectrophotometer, by Tecan (2 mentions)
Wallac, by PerkinElmer (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Rpmi medium, by Thermo Fisher Scientific (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
170 protocols using mtt dye
1
Cytotoxicity of Lactobacillus LCFS on PDLSCs
2
MTT Assay for Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) test was performed following a standard protocol [48 (link)] to exclude cytotoxic effects. Briefly, after stimulations, both cell types were incubated with 1% of MTT dye (Merck Life Science, Milano, Italy) in DMEM white for 2 h at 37 °C in an incubator, and then purple formazan crystals were dissolved in an equal volume of MTT solubilization solution. Cell viability was determined by measuring the absorbance at 570 nm with correction at 690 nm, through a spectrometer (VICTOR × 4 Multilabel Plate Reader, PerkinElmer, Waltham, MA, USA), and calculated by comparing results to control cells without any stimulus (baseline 0%).
3
Cell Proliferation Assay for Transfected Prostate Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An MTT assay was also performed in order to evaluate changes in cell proliferation. A total of 5000 transfected DU145 and PC‐3 cells were seeded onto 96‐well plates at a final volume of 100 μL medium/well (RPMI‐1640 medium containing 10% fetal bovine serum [GE Healthcare Life Sciences]). At each time point (including 0, 24, 48, 72, and 96 h), cells were stained with MTT dye (0.5 mg/mL Sigma‐Aldrich; Merck KGaA; St. Louis, MO) for 4 h at 37°C, followed by the removal of the culture medium and addition of 150 μL dimethyl sulfoxide (Sigma‐Aldrich; Merck KGaA). Then, the plates were monitored using a PowerWave XS Microplate reader (BioTek Instruments, Inc., Winooski, VT), which measured absorbance at 490 nm. The absorbance at 570 nm was used as a reference. Each experiment was performed at least in triplicate.
4
Targeted Cancer Nanotherapeutics Development
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anhydrous DTX was purchased from Jiangsu Yew (Jiangsu, China). Cetyltrimethylammonium bromide (CTAB, 98%), 3-aminopropyl triethoxysilane (APTS), mesitylene, hydroxybenzotriazole (HOBT), folic acid, methionine, fluorescein isothiocyanate (FITC) and MTT dye were purchased from Merck (Darmstadt, Germany). Tetraethyl orthosilicate (TEOS, 99%), 1-ethyl-3-[3-(dimethylamino)-propyl] carbodiimide (EDC), N-hydroxysuccinimide sodium salt (NHS) and 3a, 4, 5, 7a-tetrahydro-7-methyl-5-(tetrahydro-2, 5-dioxo-3-furanyl)-1, 3-isobenzofurandione (Epiclon B-4400) were purchased from Sigma-Aldrich Co. (St Louis, MO, USA). Dulbecco’s Modified Eagle’s Medium (DMEM) with high glucose, RPMI 1640, fetal bovine serum (FBS), trypsin, L-glutamine, penicillin and streptomycin were purchased from Biosera (Vienna, Austria). Ultra-purified water was used throughout the analyses, and all other chemicals were of analytical grades. Human breast cancer cell line (MCF-7), rat breast cancer cell line (4T1) and BALB/c mice were obtained from Pasteur Institute of Iran (Tehran, Iran).
5
Dexamethasone-Induced Cell Viability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FLSs (~5000 cells) were cultured in 96-well plates for 24 h after the addition of the indicated reagents and medium for 48 h with 100 ng/mL Dex. The cells were treated with 100 ng/mL Dex for 6, 12, 24, 36, and 48 h. Tetrazolium bromide (2 mg; MTT, Merck, Burlington, MA, USA) dye was mixed with 1 mL of PBS, and the cells were treated with 100 μL of MTT dye and incubated for 2 h in the dark. The incubation medium was carefully aspirated from the plates. A total of 100 µL of DMSO was added to the plates, and the absorbance was measured at 570 nm using a fluorescence microplate reader (VICTOR X3, PerkinElmer, Waltham, MA, USA).
6
Boron Carbide Cytotoxicity on Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bone marrow-derived macrophages (M0, M1 and M2) were placed in 96-well plates at a density of 0.15 × 105 cells/well. After 24 h, boron carbide preparations (B4C 1 and B4C 2) were added at concentrations in the range from 0.1 to 200 µg/ml and incubated for 72 h. Next, cells were incubated with MTT dye (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; 5 mg/ml)(Sigma-Aldrich) for 4 h and then lysed overnight in lysis buffer (N,N-dimethylmethanamide, sodium dodecyl sulfate, and water) at 37 °C. The absorbance of the solubilized crystal of formazan was measured at 570 nm using a Thermo Labsystems Multiskan RC microplate reader (Thermo Fisher Scientific Inc.) with Genesis Lite 3.05 Software (Thermo Life Sciences). The half maximal inhibitory concentration (IC50) value was calculated using GraphPad Prism 8 software (GraphPad Software). MTT cell viability assay for lines: RAW264.7, J774A.1 and JAWS II was performed and described in our previous work by Kozień et al. [18 (link)]. MTT assays were performed in two independent experiments, each in triplicate. The IC50 was calculated for each experiment individually and averaged on the graph for two independent experiments.
7
Synergistic Effects of CDDP and Perhexiline
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To evaluate the effects of CDDP and perhexiline combination treatment, NKX2–8+/− and NKX2–8+/+ cells were incubated with different ratios of CDDP and perhexiline and cell viability was analyzed using MTT assays. Briefly, 3000 cells/well were seeded into 96-well plates, grown for 24 h, and treated with CDDP and perhexiline alone, different weight ratios of CDDP and perhexiline combination (CDDP/perhexiline = 10:1, 5:1, 2:1, 1:1, 1:2, 1:5, and 1:10). After incubation for 24 h, 100 ml sterile MTT dye (0.5 mg/ml, Sigma) was added to each well for 4 h at 37 °C. Then, the media were removed and 150 μl dimethyl sulfoxide (DMSO, Sigma) was added. Finally, the absorbance of each well was measured at 570 nm using an EPICS XL flow cytometer (Beckman-Coulter).To evaluate the synergistic effects of CDDP and perhexiline combinations, Synergistic drug interactions were analyzed using the Chou and Talalay method [27 (link)] . The combination index (CI) for drug combinations is derived according to the equation below where n = number of drugs
According to the equation, CI <1 indicates synergy, CI = 1 indicates additivity, and CI >1 indicates antagonism. The above median effect equation and CI allow a plot of CI values at different effect levels (fas) to be determined using CalcuSyn software (Biosoft).
According to the equation, CI <1 indicates synergy, CI = 1 indicates additivity, and CI >1 indicates antagonism. The above median effect equation and CI allow a plot of CI values at different effect levels (fas) to be determined using CalcuSyn software (Biosoft).
8
MTT Assay for Cell Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An 3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide (MTT) assay was used to detect cell proliferation. Briefly, cells (1500 cells/well) were seeded in 96-well plates. After transfection, the cells were cultured for the indicated time intervals and stained with sterile MTT dye (0.5 mg/ml, Sigma). After 4 h incubation, the supernatant was aspirated, and dimethyl sulfoxide (Sigma) was added. The absorbance was measured at 490 nm. All experiments were performed in triplicate.
9
MTT Assay for Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded in 96-well plates at initial density of (0.2 × 104/well). At each time point, cells were stained with 100 μl sterile MTT dye (0.5 mg/ml, Sigma) for 4 h at 37°C, followed by removal of the culture medium and addition of 150 μl of dimethyl sulphoxide (DMSO) (Sigma, St. Louis, MO, USA). The absorbance was measured at 570 nm, with 655 nm as the reference wavelength. All experiments were performed in triplicates.
10
Glioma Cell Proliferation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An MTT assay was performed in order to determine the effect of overexpression or knockdown of JARID1B on the proliferation of glioma cells. Five thousand cells were seeded into a 96-well plate. The cells were cultured in 100 µl growth medium. At various time-points, 20 µl of sterile MTT dye (5 mg/ml; Sigma) was added, followed by incubation at 37°C for 4 h. The MTT solution was then replaced with dimethyl sulfoxide (DMSO) (200 µl) and thoroughly mixed for 30 min. The absorbance at 570 nm was measured using a microplate reader (Spectra Max 340; Molecular Devices, Sunnyvale, CA, USA) with background subtraction at 660 nm.
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!