Mtt reagent

Manufactured by Beyotime

Sourced in China

MTT reagent is a colorimetric assay used to measure cell metabolic activity. It is a yellow tetrazolium salt that is reduced by metabolically active cells, producing a purple formazan product that can be quantified spectrophotometrically.

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Lab products found in correlation

Microplate reader, by Bio-Rad (9 mentions) Dimethyl sulfoxide (dmso), by Beyotime (9 mentions) Dimethyl sulfoxide (dmso), by Merck Group (5 mentions) Microplate reader, by Thermo Fisher Scientific (4 mentions) Microplate reader, by Agilent Technologies (3 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Dimethyl sulfoxide (dmso), by Solarbio (2 mentions) Methyl thiazolyl tetrazolium (mtt), by Beyotime (2 mentions) Full wavelength multifunctional microplate reader, by Tecan (1 mentions) Crystal violet, by Beyotime (1 mentions) Paraformaldehyde (pfa), by Beyotime (1 mentions) Imark microplate reader, by Bio-Rad (1 mentions) Elx800, by Agilent Technologies (1 mentions) Facscanto 2 flow cytometry, by BD (1 mentions) Annexin 5 fitc pi apoptosis detection kit, by Vazyme (1 mentions) Enzyme labeled instrument, by Tecan (1 mentions) Gibco 10 fetal bovine serum fbs, by Thermo Fisher Scientific (1 mentions) Caco 2 human epithelial colorectal adenocarcinoma cells, by American Type Culture Collection (1 mentions) Leptomycin b, by Beyotime (1 mentions) Spectramax plus 384, by Molecular Devices (1 mentions)

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MTT assay reagent

65 protocols using mtt reagent

MTT Assay for Cell Viability

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Methylthiazolyldiphenyl tetrazolium bromide (MTT) was utilized for detecting cell viability. In brief, GI-LI-N and SK-N-BE(2) cells (100 μL) were seeded in 96-well plates at a density of 1 × 10 4 /mL and then transfected with si-XIST, si-XIST + anti-miR-653-5p, miR-653-5p, miR-653-5p + HK2, or matched controls.
MTT reagent (5mg/mL, 10 μL, Beyotime, Shanghai, China) was added to each well after transfection for 24, 48 h, or 72 h. After incubation for 4 h, the cultured medium was carefully discarded and dimethyl sulfoxide (DMSO; 150 μL) was added to per well. Lastly, the absorbance was evaluated at 490 nm under a microplate reader (Bio-Rad, Hercules, CA, USA).

Mou L., Wang L., Zhang S., & Wang Q. (2020). Long noncoding RNA XIST suppresses tumorigenesis and enhances radiosensitivity in neuroblastoma cells through regulating miR-653-5p/HK2 axis.

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Cell Viability Assay with Cytokines

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The MDA-MB-231, Hs578T and RAW264.7 cells (5×10³)/well were seeded and incubated in 96-well plates at 37°C for 24 h. Conditioned medium (CM) or drugs (IL-13, 30 ng/ml; and/or αPD-L1, 15 µg/ml) were added and the cells were further incubated at 37°C for 0, 24, 48 and 72 h. Subsequently, 100 µl MTT reagent (1 mg/ml; Beyotime Institute of Biotechnology) were added to each well and the cells were incubated at 37°C for 4 h under the same conditions. Subsequently, 100 µl DMSO (Beijing Solarbio Science & Technology) were added to each well and the optical density at 490 nm was assessed using a full-wavelength multifunctional microplate reader (Tecan, Inc.). At least five wells/group were analyzed and the experiment was repeated three times.

Meng Z., Zhang R., Wu X., Zhang M, & Jin T. (2022). PD-L1 mediates triple-negative breast cancer evolution via the regulation of TAM/M2 polarization. International Journal of Oncology, 61(6), 150.

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Cell Proliferation and Colony Formation Assay

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Transfected TPC-1 and IHH-4 cells were re-plated in 96-well plate at a density of 3,000 cells/well and cultured for 3 consecutive days for MTT assay. 10% MTT reagent (5 mg/mL; Beyotime, Shanghai, China) was added to each well and incubated for 4 h at 37°C. The MTTcontaining medium was changed to dimethylsulfoxide (150 μL; Beyotime) for 15 min-incubation. The optical density (OD) values were detected at 570 nm on a Sunrise microplate reader (TECAN, Männedorf, Switzerland). Five identical replicates were done in each transfection group.
For colony formation assay, 200 transfected TPC-1 and IHH-4 cells were re-plated in 12-well plate for another 15 consecutive days. Cell colonies were formed and stained with 0.5% crystal violet (Beyotime) for 30 min following cell fixation with 4% paraformaldehyde (Beyotime) for 30 min. Then, number of colonies was counted.

Mao Y., Huo Y., Li J., Zhao Y., Wang Y., Sun L, & Kang Z. (2021). circRPS28 (hsa_circ_0049055) is a novel contributor for papillary thyroid carcinoma by regulating cell growth and motility via functioning as ceRNA for miR-345-5p to regulate frizzled family receptor 8 (FZD8). Endocrine journal, 68(11).

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Cell Viability Assay Using MTT

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Methylthiazolyldiphenyl tetrazolium bromide (MTT) was utilized for detecting cell viability. In brief, GI-LI-N and SK-N-BE(2) cells (100 μL) were seeded in 96-well plates at a density of 1 × 10 4 /mL and then transfected with si-XIST, si-XIST + anti-miR-653-5p, miR-653-5p, miR-653-5p + HK2, or matched controls. MTT reagent (5mg/mL, 10 μL, Beyotime, Shanghai, China) was added to each well after transfection for 24, 48 h, or 72 h. After incubation for 4 h, the cultured medium was carefully discarded and dimethyl sulfoxide (DMSO; 150 μL) was added to per well. Lastly, the absorbance was evaluated at 490 nm under a microplate reader (Bio-Rad, Hercules, CA, USA).

Mu L., Wang L., Zhang S., & Wang Q. (2020). Long noncoding RNA XIST suppresses tumorigenesis and enhances radiosensitivity in neuroblastoma cells through regulating miR-653-5p/HK2 axis.

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Cell Proliferation Measurement by MTT Assay

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The cell proliferation was measured as described previously [19 (link)]. The proliferation of cells was measured by MTT assay. Briefly, MTT reagent (#ST316, Beyotime, China) was added to the cells. After 3 h incubation the supernatant was removed and 200 µL DMSO (#ST038, Beyotime, China) was added. The optical density of each well at 450 nm was detected after 2 h incubation. The experiments were repeated at least 3 independent times. 3 samples were set in each group.

Wu H., Dai R., Wang M, & Chen C. (2023). Uric acid promotes myocardial infarction injury via activating pyrin domain-containing 3 inflammasome and reactive oxygen species/transient receptor potential melastatin 2/Ca2+pathway. BMC Cardiovascular Disorders, 23, 10.

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MTT Cell Viability Assay

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In summary, transfected TE‐1 and KYSE410 were plated in a 96‐well plate for 48 h, and then hatched with 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2H‐tetrazolium bromide (MTT) reagent (20 μL, 5 mg/mL, Beyotime) for 4 h. After that, dimethyl sulfoxide (DMSO; 200 μL, Beyotime) was placed in each well after removal of the cell culture medium. The absorbance at 490 nm was then disclosed by a microplate reader (Bio‐Rad).

Chu A., Sun C., Liu Z., Liu S., Li M., Song R., Gan L., Wang Y, & Fan R. (2024). Circ‐POSTN promotes the progression and reduces radiosensitivity in esophageal cancer by regulating the miR‐876‐5p/FYN axis. Thoracic Cancer, 15(13), 1082-1094.

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Cell Proliferation Assay with MTT

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5 × 10³ cells/well treated or transfected HCC cells were cultured in 96-well plates at 37°C, 5% CO₂ for 0, 24 and 48 hours. 10 μL MTT reagent (C0009S, Beyotime, Shanghai, China) was added into each well at 37°C for another 4 hours. OD value was recorded using iMark microplate reader (Bio-Rad, Hercules, CA, USA) at an absorbance of 570 nm.

Zhang K., Fang T., Shao Y, & Wu Y. (2021). TGF-β-MTA1-SMAD7-SMAD3-SOX4-EZH2 Signaling Axis Promotes Viability, Migration, Invasion and EMT of Hepatocellular Carcinoma Cells. Cancer Management and Research, 13, 7087-7099.

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Cell Viability Assay with TG and ICA

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Various concentrations of TG and ICA were selected to treat the cells with or without PI3K/AKT inhibitor LY294002 in the 96-well plate. At 37 ^oC, the cells were cultured for 4 h with 10 μl MTT reagent (Beyotime, Shanghai, China) at 5 % CO₂ in dark. Then, 100 μl DMSO was added to dissolve the formazan in each well. The measurement of optical density (OD) was perfomed by a microplate reader (Bio-Tek, USA) at 490 nm. The OD of treated group/ the OD of control group × 100 % was calculated as cell viability.

Li H., Zhang X., Qi X., Zhu X, & Cheng L. (2019). Icariin Inhibits Endoplasmic Reticulum Stress-induced Neuronal Apoptosis after Spinal Cord Injury through Modulating the PI3K/AKT Signaling Pathway. International Journal of Biological Sciences, 15(2), 277-286.

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Cell Viability Assay using MTT

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Cell viability was determined using MTT reagent (Beyotime Institute of Biotechnology, Haimen, China) in accordance with the manufacturer’s instructions. In brief, HUVECs were grown in 96-well plates. After the indicated treatments, 10 μL of MTT solution (5 mg/mL) was added to each well, followed by incubation at 37 °C with 5% CO₂ for another 4 h. Subsequently, 200 μL DMSO was added to each well to dissolve any crystals, and the plates were agitated for 10 min. The OD values of each well were measured at the wavelength of 490 nm.

Xie Y., Lou D, & Zhang D. (2021). Melatonin Alleviates Age-Associated Endothelial Injury of Atherosclerosis via Regulating Telomere Function. Journal of Inflammation Research, 14, 6799-6812.

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MTT Assay for Hg Cytotoxicity in CEK Cells

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The CEK cells viability was quantitated using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Then the cells in 96-well plants were treated with PBS or different dosages of Hg for 24 h. In parallel, the CEK cells were exposed to the 15-μmol/L Hg for 24 h in the presence or absence of 1-μmol/L 4-PBA. Ten microliters of MTT reagent (Beyotime Institute of Biotechnology, Wuhan, China) was added to each well, and the cells were incubated for another 4h at 37°C followed by adding dimethyl sulfoxide solution to dissolve formazan crystals. Subsequently, the absorbance was determined at 570-nm wavelength by an enzyme-linked immunosorbent assay reader (Bio-tek ELX800; Winooski, VT). The calculation of cell activity was based on a previous study (Ma et al., 2018b ).

Ma Y., Shi Y., Zou X., Wu Q, & Wang J. (2020). Apoptosis induced by mercuric chloride is associated with upregulation of PERK-ATF4-CHOP pathway in chicken embryonic kidney cells. Poultry Science, 99(11), 5802-5813.

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