Mtt assay

Manufactured by Abcam

Sourced in United Kingdom, United States

The MTT assay is a colorimetric assay used to measure the activity of enzymes that reduce the tetrazolium dye MTT to its insoluble formazan, which has a purple color. This assay is commonly used to assess cell viability and proliferation.

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Lab products found in correlation

Oxaliplatin, by MedChemExpress (2 mentions) Clariostar plate reader, by BMG Labtech (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Penicillin, by Merck Group (2 mentions) Tumor necrosis factor (tnf), by Thermo Fisher Scientific (2 mentions) Mcf 7 breast cancer, by American Type Culture Collection (1 mentions) Cyquant cell proliferation assay, by Thermo Fisher Scientific (1 mentions) Spectramax m3, by Molecular Devices (1 mentions) Matrigel invasion chamber, by Corning (1 mentions) Flexstation 3, by Molecular Devices (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Streptomycin, by Merck Group (1 mentions) Mycoalert mycoplasm detection kit, by Lonza (1 mentions) Infinite m200 pro, by Tecan (1 mentions) Versamax, by Molecular Devices (1 mentions) Facscalibur, by BD (1 mentions) Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions)

Spelling variants of this product

MTT assay kit (97 citations) MTT Cell Proliferation Assay Kit (36 citations) 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay kit (3 citations) MTT colorimetric assay (2 citations) MTT) reduction assay (2 citations)

50 protocols using mtt assay

Evaluating pAp Cytotoxicity in A549 Cells

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A549 cells (2× 10³ cells/well) were seeded in 96-well plates for 24 h, and then the cells were treated with various concentrations of pAp (1 to 6 mM) (catalog no. A5763; Sigma). The viability of pAp-treated A549 cells was validated by MTT assay (Abcam) in accordance with the manufacturer’s instructions.

Liu Y.C., Mok B.W., Wang P., Kuo R.L., Chen H, & Shih S.R. (2021). Cellular 5′-3′ mRNA Exoribonuclease XRN1 Inhibits Interferon Beta Activation and Facilitates Influenza A Virus Replication. mBio, 12(4), e00945-21.

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Sorbitol Cytotoxicity in HepG2 and MCF7 Cells

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2,500 cells were seeded for 24 hours in 96-well plates. After washing the wells twice with PBS, fresh medium was added to each well with or without (control) sorbitol for 4, 8, 12, 24, or 48 more hours and cellular DNA content was measured using the CyQUANT Cell Proliferation Assay (ThermoFisher Cat. No. C7026). CyQUANT was excited at 485 nm and detected at 528 nm using a Biotek Synergy 1 Hybrid Multi-mode Microplate Reader (Biotek Instruments Inc., Winooski, VT). Cell viability was assessed with the MTT assay (Abcam Cat.No. 211091) measuring absorbance at 570 nm with the Synergy 1 plate reader. HepG2 hepatoma (catalogue number HB-8065) and MCF7 breast cancer (catalogue number HTB-22) cell lines were obtained from ATCC (Manassas, VA, USA).

Oaks Z., Patel A., Huang N., Choudhary G., Winans T., Faludi T., Krakko D., Duarte M., Lewis J., B.e.c.k.f.o.r.d., Blair S., Kelly R., Landas S., Middleton F., Asara J., Chung S., Fernandez D., Banki K, & Perl A. (2023). CYTOSOLIC ALDOSE METABOLISM CONTRIBUTES TO PROGRESSION FROM CIRRHOSIS TO HEPATOCARCINOGENESIS. Nature metabolism, 5(1), 41-60.

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Metabolic Activity of ALI-Cultured Tissue Slices

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The overall metabolic activity of the ALI-cultured slices was investigated by methyl thiazolyl tetrazolium (MTT) assay (Abcam) at D0 (fresh), D1, D3, D5, D7 and D10 of cultivation. According to manufacturer’s instructions, the basic culture medium was exchanged with 1 ml MTT-containing culture medium (1:100 dilution). Following 4 h incubation at 37

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C and 5% CO2, the slices were then directly placed in 600 μl MTT solvent reagent and left for incubation at 37

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C and 5% CO2 overnight. The absorbance of supernatant was measured at 570 nm in a microtiter plate reader. The average values from triplicate readings per probe were adjusted by subtracting the average value for the blank value (MTT solvent reagent). The weight of wipe-dried tissue slice was determined using an analytical balance. The final metabolic activity value is given as measured optical density (OD) per weight unit (gram). Overall, 24 slices from 7 different patients were analyzed (4 slices per time point).

Chabanovska O., Lemcke H., Lang H., Vollmar B., Dohmen P.M., David R., Etz C, & Neßelmann C. (2023). Sarcomeric network analysis of ex vivo cultivated human atrial appendage tissue using super-resolution microscopy. Scientific Reports, 13, 13041.

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MTT Assay for A549 Cell Viability

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The viability of A549 cells was measured using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay(Abcam, Cambridge, UK). Human lung epithelial A549 cells were used to analyze interactions with B. cepaciaOMVs, as epithelial cells represent the first line of defense against bacteria or bacterial products. Cells were seeded at a concentration of 2.0 × 10⁴/well in a 96-well microplate. After treatment with different concentrations of B. cepacia OMVs for 24 h, cell viability was measured 2 h after treatment with MTT reagent at 590 nm. Cell viability was calculated as follows: cell viability (%) = the absorbance of OMV-treated cells/the absorbance of the control cells × 100%.

Kim S.Y., Kim M.H., Son J.H., Kim S.I., Yun S.H., Kim K., Kim S., Shin M, & Lee J.C. (2020). Outer membrane vesicles produced by Burkholderia cepacia cultured with subinhibitory concentrations of ceftazidime enhance pro-inflammatory responses. Virulence, 11(1), 995-1005.

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Doxycycline-Induced Cell Viability Assay

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293T-S cells and 293T-S-ACE2 cells seeded in clear 96-well plates at different densities were induced with 1 μg/ml doxycycline. Cell viability was measured using an MTT assay (Abcam) at different times after induction, according to the manufacturer’s protocol.

Nguyen H.T., Zhang S., Wang Q., Anang S., Wang J., Ding H., Kappes J.C, & Sodroski J. (2021). Spike Glycoprotein and Host Cell Determinants of SARS-CoV-2 Entry and Cytopathic Effects. Journal of Virology, 95(5), e02304-20.

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Chondrocyte Proliferation Assay

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Human articular chondrocytes were seeded (2,500 cells in each well of 96-well-plate) and incubated in media as listed in Table 1. No-EV was used as the control. Cells were incubated at 37°C and 5% CO₂ for 48 hours to proliferate. The cell proliferation rate was assessed by performing a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay (Abcam, Cambridge, UK) following the manufacturer’s protocols. The proliferation rate was equivalent to the relative absorbance measured at 562 nm (SpectraMax M3, Molecular Devices, California, USA) at time points of 0 hours (as used for normalization) and 48 hours. The proliferation rate was calculated based on the OD values obtained from two time points.

Nguyen T.H., Dao H.H., Duong C.M., Nguyen X.H., Hoang D.H., Do X.H., Truong T.Q., Nguyen T.D., Nguyen L.T, & Than U.T. (2022). Cytokine-primed umbilical cord mesenchymal stem cells enhanced therapeutic effects of extracellular vesicles on osteoarthritic chondrocytes. Frontiers in Immunology, 13, 1041592.

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Oxaliplatin Cytotoxicity in Colorectal Cell Lines

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SW620, SW620 OxR, HCT116, HCT116 OxR, HT29, HT29 OxR, SW480, and SW480 OxR cell lines were plated into tissue culture 96-well black-walled plates at a concentration of 3000 cells/well and incubated overnight at 37°C. A 10 mM stock oxaliplatin suspension was created by dissolving oxaliplatin (MedChemExpress) in molecular grade water via sonication and heating. Cell culture media was replaced with oxaliplatin treatments ranging from 0 to 1000 µM and incubated for 72 hr. Following treatments, an MTT assay (Abcam) was carried out according to the manufacturer’s protocol. The plates were then read using a plate reader (BioTek µQuant) at 590 nm absorbance using gen5 software. Control wells containing the MTT solution without cells were used for background subtraction.

Greenlee J.D., Lopez-Cavestany M., Ortiz-Otero N., Liu K., Subramanian T., Cagir B, & King M.R. (2021). Oxaliplatin resistance in colorectal cancer enhances TRAIL sensitivity via death receptor 4 upregulation and lipid raft localization. eLife, 10, e67750.

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MTT Assay for LSD1 Inhibitor Cytotoxicity

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An MTT assay (Abcam) was carried out to measure the effects of LSD1 inhibitors on cell viability and proliferation. Cells were plated in 96-well plates for 24 h prior to addition of inhibitors (1.0 × 10⁴ cells/well). The indicated concentrations of SP-2509 or OG-L002 were added and cells were incubated at 37°C for an additional 24 h before conducting the MTT assay according to the manufacturer’s protocol. Percent cytotoxicity was determined:

% Cytotoxicity = \frac{(100 x (Control - Sample))}{Control}

Harancher M.R., Packard J.E., Cowan S.P., DeLuca N.A, & Dembowski J.A. (2020). Antiviral Properties of the LSD1 Inhibitor SP-2509. Journal of Virology, 94(19), e00974-20.

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Evaluating U937 Cell Viability

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U937 cells were seeded in a 96-well plate at a concentration of 0.2 × 10⁶ cells/ml, 18 hours before compound addition, which was performed in triplicate, with all experiments including a final concentration of 0.1% DMSO (v/v). To quantify U937 cell viability, metabolic activity was assessed 72 hours after compound exposure using an MTT assay (Abcam), as described previously (104 (link)). Briefly, thiazolyl blue tetrazolium bromide was dissolved in PBS and added to cells at a final concentration of 0.25 mg/ml and incubated at 37°C for 3 hours. The reaction was stopped by the addition of 50 μl of acidified 10% SDS, followed by reading of absorbance at 570 nm. Viability was defined relative to DMSO-containing controls incubated for the same period of time.

Foulkes D.M., Byrne D.P., Yeung W., Shrestha S., Bailey F.P., Ferries S., Eyers C.E., Keeshan K., Wells C., Drewry D.H., Zuercher W.J., Kannan N, & Eyers P.A. (2018). Covalent inhibitors of EGFR family protein kinases induce degradation of human Tribbles 2 (TRIB2) pseudokinase in cancer cells. Science signaling, 11(549), eaat7951.

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Cytotoxicity Evaluation of Extracted Samples

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The cytotoxicity of the extracted samples was quantified using the MTT assay (Abcam). In total, 5x10³ cells/well were cultured in 96-well plates for 24 h at 37˚C with 5% CO₂, before fresh medium containing serial concentrations of the sample extracts were added. After 72 h of incubation at 37˚C with 5% CO₂, the extract medium was removed and MTT 5 mg/ml dissolved in fresh media was added into each well, which was then incubated for 3 h at 37˚C. This solution was then removed and DMSO was added for incubation for 5 min at 27˚C. The absorbance was measured using a spectrophotometer at wavelengths of 492 with a reference wavelength of 630 nm use for all samples (30 (link)). The absorbance data were calculated by subtracting the 492 nm result from the 630 nm result, which was subsequently provided as % viability compared with the untreated control. The IC₅₀ was reported as the concentration of the extracts that inhibited cell proliferation by 50%. IC₅₀ was determined using curves constructed by plotting cell survival (%) against the concentration of extracted samples (31 (link),32 ). The results of MTT assay (IC₅₀-dependent manner) were used for further assays for cell proliferation and apoptosis.

Syukriya A.J., Bankeeree W., Prasongsuk S, & Yanatatsaneejit P. (2023). In vitro antioxidant and anticancer activities of Smilax corbularia extract combined with Phellinus linteus extract against breast cancer cell lines. Biomedical Reports, 19(3), 63.

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