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Mts assay

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The MTS assay is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity experiments. The assay involves the chemical reduction of a tetrazolium compound, MTS, to a colored formazan product that is soluble in tissue culture medium. The amount of formazan produced is directly proportional to the number of living cells in culture.

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Lab products found in correlation

Mts reagent, by Abcam (3 mentions) Bca kit, by Thermo Fisher Scientific (3 mentions) Trizol, by Thermo Fisher Scientific (2 mentions) Prism 7, by GraphPad (1 mentions) Bay11 7085, by Apexbio (1 mentions) Erlotinib, by Cayman Chemical (1 mentions) Primescript rt reagent kit, by Takara Bio (1 mentions) Milli q grade water, by Merck Group (1 mentions) Sybr premix ex taqtm 2 tli rnaseh plus, by Takara Bio (1 mentions) Compusyn software, by ComboSyn (1 mentions) Htx multi mode microplate reader, by Synergy Software (1 mentions) Cnt 57, by CELLnTEC (1 mentions) Amiloride, by Merck Group (1 mentions) Rifampin, by Merck Group (1 mentions) Fk506, by Merck Group (1 mentions) Apilimod, by Merck Group (1 mentions) Torin1, by Merck Group (1 mentions) E coli lps, by Merck Group (1 mentions) Bafilomycin a1, by Merck Group (1 mentions) Thapsigargin, by Merck Group (1 mentions)

43 protocols using mts assay

1

MTS Assay for Cell Viability

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The MTS assay (Biovision) was employed to determine cell viability following the manufacturer’s protocol. 1000/well cells were seeded in 96-well plate. 10 μl /well MTS Reagent was added into each well and incubated for 1 h at each time point. Absorbance was measured at 490 nm using SpectraMax M3 reader with SoftMax Pro 6 software for data acquisition and analysis. For cells proliferation assay, results of each group were normalized to Day 0. For IC50 determination of ATM inhibitor and Enzalutamide, cells were treated by a series of concentration for 72 h. IC50 value was calculated by GraphPad Prism software. All assays were performed in triplicate.
Yin Y., Xu L., Chang Y., Zeng T., Chen X., Wang A., Groth J., Foo W.C., Liang C., Hu H, & Huang J. (2019). N-Myc promotes therapeutic resistance development of neuroendocrine prostate cancer by differentially regulating miR-421/ATM pathway. Molecular Cancer, 18, 11.
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2

Micelle Cytotoxicity Evaluation in Diverse Cell Lines

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The in vitro cytotoxicity of micelles was evaluated with the (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) assay (Biovision, Milpitas, CA, USA). B16F10, PC3, 22Rv1, NCI-H460, WEHI-274.1, and RWPE-1 cells were plated at a density of 2000 cells/well in 96-well plates. 0.5–50 μM of KLAK-MCP-1, KLAK-scr-MCP-1, KLAK, MCP-1, scr-MCP-1, and empty micelles were incubated with cells for 72 hours before addition of the MTS reagent and evaluation using a Varioskan Lux microplate reader at an absorbance wavelength of 490 nm. After subtracting the background absorbance of culture media + MTS reagent alone, cell viability data was normalized to PBS-treated cells, and IC50 values were calculated using GraphPad Prism 7 (GraphPad, San Diego, CA, USA). For each assay, n = 6 was used for each concentration.
Trac N., Chen L.Y., Zhang A., Liao C.P., Poon C., Wang J., Ando Y., Joo J., Garri C., Shen K., Kani K., Gross M.E, & Chung E.J. (2020). CCR2-targeted micelles for anti-cancer peptide delivery and immune stimulation. Journal of controlled release : official journal of the Controlled Release Society, 329, 614-623.
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3

Quantifying Cell Metabolic Activity in Scaffolds

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The metabolic activity of cells in scaffolds was quantitatively evaluated by mitochondrial dehydrogenase activity using MTS assay (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) from BioVision (Catalog #K300-500), according to the manufacturer’s protocol. Scaffolds were located in 48-well plates. Cells were seeded onto the scaffolds (0.75 × 104 cells in 400 mL per well) as per the ratio of 10% MTS assay into cell-scaffold media and incubated for 4 h. The cell scaffolds were cultured from days 3, 5, 10, and 30 at 37 °C/5% CO2. After incubation for an hour, the cultures were measured at 490 nm [27 (link)].
Ebrahimi S., Hanim Y.U., Sipaut C.S., Jan N.B., Arshad S.E, & How S.E. (2021). Fabrication of Hydroxyapatite with Bioglass Nanocomposite for Human Wharton’s-Jelly-Derived Mesenchymal Stem Cell Growing Substrate. International Journal of Molecular Sciences, 22(17), 9637.
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4

Cytotoxicity Evaluation of Magnetic Nanoparticles

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hAECs were plated in 96-well plates at a cell density of 2000 cells/well and varying concentrations of CREKA peptide, CREKA-Ms, Fe-Ms, and Mn-Ms in PBS were added to wells. The plates were incubated at 37 °C and 5% CO2 for 72 h, and cell viability was determined via MTS assay (BioVision, Milpitas, CA, USA) according to manufacturer’s instructions. Absorbance at 490 nm was used to measure the enzymatic activity level, and 100% viability using non-treated, healthy cells was used as a positive control. Viability of each treatment group was measured from six replicates.
Poon C., Gallo J., Joo J., Chang T., Bañobre-López M, & Chung E.J. (2018). Hybrid, metal oxide-peptide amphiphile micelles for molecular magnetic resonance imaging of atherosclerosis. Journal of Nanobiotechnology, 16, 92.
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5

Cell Viability and Proliferation Assay

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The MTS assay (Biovision, San Francisco Bay, CA, USA) was employed to determine cell viability following the manufacturer’s protocol. Briefly, 1000/well cells were seeded in a 96-well plate. Ten μL/well MTS Reagent was added into each well and incubated for at least 1 h. Absorbance was measured at 490 nm using the SpectraMax M3 reader with SoftMax Pro 6 software (V5.3) for data acquisition and analysis. For cells proliferation assay, the results of each group were normalized to Day 0. For IC50 determination of CPT1-inhibitor, cells were treated by a series of concentration for 72 h. IC50 value was calculated by the GraphPad Prism software (San Diego, CA, USA). All assays were performed in triplicate.
Xu H., Li S., Sun Y., Xu L., Hong X., Wang Z, & Hu H. (2021). ELOVL5-Mediated Long Chain Fatty Acid Elongation Contributes to Enzalutamide Resistance of Prostate Cancer. Cancers, 13(16), 3957.
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6

Cytotoxicity Evaluation of MCP-1 Targeting Agents

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In a 96-well plate, WEHI 274.1 murine monocytes were seeded at a density of 4000 cells/well in supplemented DMEM and incubated for 24 h. Different concentrations of MCP-1 PAMs, scrambled PAMs, MCP-1 peptides, scrambled peptides, and DSPE-PEG(2000)-methoxy in 10 μL of PBS were added to the wells. After 24 or 72 h, cell viability was determined by a (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) MTS assay (BioVision, Milpitas, CA), according to the manufacturer’s instructions. IC50 values were determined from the best fit of the cell viability (%) versus concentration (μM) plot. In the analysis, 100% viability using nontreated, healthy cells was used as a positive control and to normalize the data. For each assay, 6 different wells per concentration per treatment group were assessed (n = 6).
In vitro cell viability of 22Rv1 and PC3 prostate cancer cells were performed as described above, except they were seeded at 2000 cells/well in supplemented RPMI 1640 media.
Poon C., Chowdhuri S., Kuo C.H., Fang Y., Alenghat F.J., Hyatt D., Kani K., Gross M.E, & Chung E.J. (2017). Protein Mimetic and Anticancer Properties of Monocyte-Targeting Peptide Amphiphile Micelles. ACS biomaterials science & engineering, 3(12), 3273-3282.
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7

Optimizing Cell Viability Analysis

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The MTS assay (Biovision) was employed to determine cell viability. 1000/well cells were seeded in 96-well plate. 10 μl /well MTS Reagent was added into each well and incubated for 1 hour at each time point. Absorbance was measured at 490nm using SpectraMax M3 reader with SoftMax Pro 6 software for data acquisition and analysis. For cells treated with siRNAs, 5 pmol siLDHA or siCtrl was transfected into each well and cell viability was measured at each time point. For cells treated with FX11 (Millopore), 3 μM FX11 was added in culture medium followed by cell viability measurement. For IC50 determination of FX11, cells were treated by a series of concentration for 72 hours. IC50 value was calculated by GraphPad Prism software.
Xu L., Ma E., Zeng T., Zhao R., Tao Y., Chen X., Groth J., Liang C., Hu H, & Huang J. (2019). ATM Deficiency Promotes Progression of CRPC by Enhancing Warburg Effect. Endocrine-related cancer, 26(1), 59-71.
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8

Evaluating Gefitinib, Erlotinib, and Cetuximab Efficacy

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Cells were seeded at a density of 10,000 cells/well in a 96-well plate and allowed to attach for eight hours prior to drug treatment. For viability testing, cells were treated with 5 μM gefitinib (#076091; Matrix Scientific, Columbia, SC), 10 μM erlotinib (#10483; Cayman Chemical; Ann Arbor, MI), or 10 nM cetuximab (#NBP2-75903; Novus Biologicals; Centennial, CO), with or without BAY 11–7085 (#B3033; ApexBio Technology; Houston, TX) at 2 μM or 3.3 μM for 48 hours. DMSO vehicle controls were included. For all treatments the final DMSO concentration was 1%. After 48 hours cell viability was assessed using the MTS assay (Biovision; Milpitas, CA) as previously described [35 (link)]. Viability was calculated by normalizing each cell line to vehicle treatment alone.
Landmesser M.E., Raup-Konsavage W.M., Lehman H.L, & Stairs D.B. (2020). Loss of p120ctn causes EGFR-targeted therapy resistance and failure. PLoS ONE, 15(10), e0241299.
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9

Evaluating PAM Compatibility with Monocytes

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To determine the compatibility of PAMs with monocytes, 1 µM, 10 µM, and 100 µM of S-MCP-1 PAMs, S-Scrambled PAMs, C-MCP-1 PAMs, C-Scrambled PAMs, MCP-1 peptides, and scrambled MCP-1 peptides were incubated with WEHI-274.1 monocytes (4000 cells/well). Cell viability was determined at 24 h using a (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) MTS Assay (BioVision, Milpitas, CA, USA). The IC50 values were measured by analyzing cell viability % vs. concentration, and the 100% viability represented non-treated monocytes (positive control).
Joo J., Poon C., Yoo S.P, & Chung E.J. (2018). Shape Effects of Peptide Amphiphile Micelles for Targeting Monocytes. Molecules, 23(11), 2786.
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10

FMDV Vaccine Strain Propagation and Characterization

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Baby Hamster Syrian Kidney (BHK-21cells) were cultured in Dulbeccoʼs modified Eagleʼs medium (DMEM) supplemented with 10% FBS. FMDV strains, O/MYA98/BY/2010 and Asia1/JSL/ZK/06 investigated extensively as vaccine strains, were provided by Zhongnongweite biotechnology Co., Ltd. Each strain was propagated in BHK-21 cells at a multiplicity of infection of 0.001 and incubated in 5% CO2 at 37 °C. A PrimeScriptTM RT reagent kit containing gDNA Eraser and SYBR Premix Ex TaqTM II (Tli RNaseH Plus) was purchased from TaKaRa (Dalian, China). MTS assay was available from Abcam (Cambridge, UK). The 50% tissue culture infectious dose (TCID50) was measured with the Reed and Muench method. Trehalose, CuSO4·5H2O were purchased from sigma (MO, USA). All other chemicals were analytical grade reagents purchased from Beijing Chemical works (Beijing, China) and all solutions were prepared using Milli-Q grade water (Millipore,USA).
Li J., Chang Y., Yang S., Zhou G, & Wei Y. (2022). Formulation enhanced the stability of Foot-and-mouth virus and prolonged vaccine storage. Virology Journal, 19, 207.
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