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Mtt labeling reagent

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The MTT labeling reagent is a colorimetric assay used to measure cell metabolic activity. It is a widely-used tool for evaluating cell proliferation, cytotoxicity, and viability in various cell-based applications.

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Lab products found in correlation

Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Flutamide, by Merck Group (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions) Mars software, by BMG Labtech (1 mentions) Clariostar reader, by BMG Labtech (1 mentions) Cell proliferation kit 1 mtt, by Merck Group (1 mentions) Mdck cells, by American Type Culture Collection (1 mentions) Glomax discover microplate reader, by Promega (1 mentions) Eon microplate spectrophotometer, by Agilent Technologies (1 mentions) Elx800 microplate photometer, by Agilent Technologies (1 mentions) Synergy h1 hybrid reader, by Agilent Technologies (1 mentions) Imark microplate reader, by Bio-Rad (1 mentions)

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MTT reagent (974 citations) ) labeling reagent (2 citations)

13 protocols using mtt labeling reagent

1

MTT Assay for Cell Proliferation

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Cell proliferation kit was following the manufacturer’s instructions and previously described methods (Tsai et al., 2016 (link)). In brief, 1×104 of 293T cells were cultured in 96-well plates. After 2BP treatment at 37°C for 24 h, the MTT labeling reagent (Sigma-Aldrich, Darmstadt, Germany) was added to each well at a final concentration of 0.5 mg/mL for additional 4 h incubation. Finally, the reaction was terminated by adding the stop solution, and the absorbance was measured at 570 nm.
Tien C.F., Tsai W.T., Chen C.H., Chou H.J., Zhang M.M., Lin J.J., Lin E.J., Dai S.S., Ping Y.H., Yu C.Y., Kuo Y.P., Tsai W.H., Chen H.W, & Yu G.Y. (2022). Glycosylation and S-palmitoylation regulate SARS-CoV-2 spike protein intracellular trafficking. iScience, 25(8), 104709.
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2

Evaluating Flutamide-Induced Hepatotoxicity in HepG2 Cells

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Cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (Sigma-Aldrich, St. Louis, MO) assay. HepG2 cells have been widely used for the detection of flutamide-dependent liver injury in previous studies [27 –29 (link)]. In the present study, HepG2 cells were seeded at a density of 2 × 104 per well in 96-well plates. MiRNA transfection and lansoprazole induction were carried out as described above. After a period of 12 h incubation, flutamide (Sigma-Aldrich, St. Louis, MO) was dissolved in ethanol and added at a final concentration of 1 mM [29 (link),30 (link)] to each well and incubated at 37 °C, 5% CO2 for 4 h. MTT labeling reagent (Sigma-Aldrich) was added to the cells, and the cells were incubated at 37 °C for 2 h. Dimethyl sulfoxide (DMSO, Sigma-Aldrich, St. Louis, MO) was then added, and the absorbance of stained cells was measured at 570 nm. Five experiments were performed independently.
Chen Y., Zeng L., Wang Y., Tolleson W.H., Knox B., Chen S., Ren Z., Guo L., Mei N., Qian F., Huang K., Liu D., Tong W., Yu D, & Ning B. (2017). The expression, induction and pharmacological activity of CYP1A2 are post-transcriptionally regulated by microRNA hsa-miR-132-5p. Biochemical pharmacology, 145, 178-191.
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3

MTT Assay for Cell Viability Assessment

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For assessing the cell viability, cells were seeded in a 96-well plate (at concentrations of 2 × 103/0.32 cm2 and 4 × 103/0.32 cm2). Then, 24 h, 48 h, and 72 h after irradiation, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) labeling reagent (Sigma-Aldrich, St Louis, MO, USA) (final concentration 0.5 mg/mL) was added into each well. Two hours after incubation in a humidified atmosphere of 5% CO2 at 37 ◦C, MTT solution was removed and 150 µL of DMSO was added into each well to extract the blue MTT–formazan crystals. The absorbance measurement was performed at a 570 nm wavelength using a CLARIOstar reader (BMG LABTECH, Ortenberg, Germany). The collected data were analyzed using MARS software (BMG LABTECH, Ortenberg, Germany).
Alhaddad L., Chuprov-Netochin R., Pustovalova M., Osipov A.N, & Leonov S. (2023). Polyploid/Multinucleated Giant and Slow-Cycling Cancer Cell Enrichment in Response to X-ray Irradiation of Human Glioblastoma Multiforme Cells Differing in Radioresistance and TP53/PTEN Status. International Journal of Molecular Sciences, 24(2), 1228.
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4

MTT Assay for Cell Viability

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Cells were grown in 98-well culture plates, and incubated with 100 μL/well of the MTT labeling reagent (Sigma-Aldrich) for 3 h. Blue formazan crystals were solubilized with acidified isopropanol, and formazan levels were determined at 570 nm.
Shin S.H., Lee G.Y., Lee M., Kang J., Shin H.W., Chun Y.S, & Park J.W. (2018). Aberrant expression of CITED2 promotes prostate cancer metastasis by activating the nucleolin-AKT pathway. Nature Communications, 9, 4113.
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5

Cytotoxicity Evaluation of Nanoparticles

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Cells were cultured in a 96-well microplate overnight and then incubated with nanoparticles. Next, the MTT labeling reagent (Sigma) was added and the cells were incubated in a humidified atmosphere for 4 h. Solubilization solution is then added followed by further incubation in a humidified atmosphere overnight. Finally, the microplate was evaluated with the use of an ELISA reader at 550– 600 nm with a reference wavelength of >650 nm.
Liang L., Liu Z, & Barman I. (2019). Decoding Live Cell Interactions with Multi-Nanoparticle Systems: Differential Implications for Uptake, Trafficking and Gene Regulation. ACS applied materials & interfaces, 11(37), 33659-33666.
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6

Cytotoxicity Assay with Doxorubicin

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Cells were grown in 98-well culture plates. After being treated with doxorubicin, H2O2 and PEITC for 24 h, the cells were incubated with 100 μl of MTT labeling reagent (Sigma–Aldrich) for 3 h. Blue formazan crystals were solubilized with acidified isopropanol, and formazan levels were determined at 570 nm.
Shin S.H., Yoon H., Chun Y.S., Shin H.W., Lee M.N., Oh G.T, & Park J.W. (2014). Arrest defective 1 regulates the oxidative stress response in human cells and mice by acetylating methionine sulfoxide reductase A. Cell Death & Disease, 5(10), e1490-.
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7

Cytotoxicity Assay for SARS-CoV-2 Host Cells

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Drug cytotoxicity on A549(ACE2) cells and Vero E6 cells were quantify by propidium iodide staining and flow cytometry as described (34 (link)). Drug toxicity on MDCK cells was quantified using Cell Proliferation Kit I (MTT) (Sigma) and the protocol suggested by the manufacturer. Briefly, MDCK cells (ATCC) were seeded into a 12 well plate at 1 × 105 cells per well. Cells were cultured overnight, and then treated with RDS for one day, and then cultured in the medium supplemented with MTT labeling reagent (Sigma). Cells were incubated with the labeling reagent for 4 h, followed by the addition of MTT solubilization solution. The plate was incubated overnight, and then the absorbance was measured using GloMax Discover Microplate Reader (Promega).
Hetrick B., Yu D., Olanrewaju A.A., Chilin L.D., He S., Dabbagh D., Alluhaibi G., Ma Y.C., Hofmann L.A., Hakami R.M, & Wu Y. (2021). A traditional medicine, respiratory detox shot (RDS), inhibits the infection of SARS-CoV, SARS-CoV-2, and the influenza A virus in vitro. Cell & Bioscience, 11, 100.
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8

Quantifying Cytotoxicity via MTT Assay

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An MTT labeling reagent (Sigma-Aldrich, St Louis, MI) was used to evaluate cytotoxicity. Cells were plated in 24-well dishes and cultured in 0.5 ml of medium per well. After cells were treated with each compound for 24 hours, cells were incubated with 5 mg·mL−1 of MTT for 3 hours. Blue formazan crystals were solubilized with acidified isopropanol, and formazan levels were determined at 570 nm using a spectrophotometer.
Choi Y.J., Shin H.W., Chun Y.S., Leutou A.S., Son B.W, & Park J.W. (2016). Diacetoxyscirpenol as a new anticancer agent to target hypoxia-inducible factor 1. Oncotarget, 7(38), 62107-62122.
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9

MTT Assay for Cell Viability

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MDA-MB-231 and MDA-MB-468 cell viabilities were evaluated with an MTT assay modified from the MTT Assay Protocol from Millipore Sigma. Cells were seeded at a density of about 80% confluency in 96-well plate. After the incubation with treatments, 10 μL of the MTT labeling reagent (final concentration of 0.5 mg/mL, 475989, Sigma-Aldrich) was added to each well and incubated for 4 h in a cell incubator. The formazan crystals were dissolved in lysis solution (10% NP-40, 10 mM HCl) overnight at 37 °C, and absorbance was measured at 570 nm with correction at 690 nm with a BioTek Eon Microplate Spectrophotometer.
Guo Z., Bergeron K.F, & Mounier C. (2024). Oleate Promotes Triple-Negative Breast Cancer Cell Migration by Enhancing Filopodia Formation through a PLD/Cdc42-Dependent Pathway. International Journal of Molecular Sciences, 25(7), 3956.
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10

MTT Cell Proliferation Assay

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Cell proliferation was assessed using MTT kit. The cells were seeded on 96-well microplates at a density of 3000 cells per well. At 1–3 days, the cells were incubated with 20 μl of MTT labeling reagent (0.5 mg/ml, Sigma-Aldrich) for 4 h, followed by the addition of 100 μl of dimethylsulfoxide (Sigma-Aldrich) into each well. The plates were kept in darkroom for 15 min and OD value was measured with BioTek ELx800 microplate photometer (BioTek ELx800, SN 21 18 05, US). Investigated wavelength: 570 nm, reference wavelength: 690 nm.
Peng H., Gong P.G., Li J.B., Cai L.M., Yang L., Liu Y.Y., Yao K.T, & Li X. (2016). The important role of the receptor for activated C kinase 1 (RACK1) in nasopharyngeal carcinoma progression. Journal of Translational Medicine, 14, 131.
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