Mtt cell viability assay kit

Manufactured by Biotium

Sourced in United States

The MTT Cell Viability Assay Kit is a colorimetric assay used to measure the metabolic activity of cells. The assay is based on the reduction of the tetrazolium salt MTT to a colored formazan product by the mitochondria of viable cells. The amount of formazan produced is directly proportional to the number of living cells, which can be quantified by measuring the absorbance at a specific wavelength.

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Lab products found in correlation

Imark microplate reader, by Bio-Rad (4 mentions) Gene pulser mxcell system, by Bio-Rad (2 mentions) Hiperfect reagent, by Qiagen (2 mentions) Silencer sirna, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Escherichia coli serotype 0111 b4, by Merck Group (1 mentions) Fluostar omega, by BMG Labtech (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Prism 9, by GraphPad (1 mentions) Dimethyl sulfoxide (dmso), by Corning (1 mentions) Heparinase, by Merck Group (1 mentions) P24 elisa, by PerkinElmer (1 mentions) Keratinocyte growth medium, by Lonza (1 mentions) Infinite 200, by Tecan (1 mentions) Bafilomycin, by Merck Group (1 mentions) Bafilomycin a1, by Merck Group (1 mentions) Letrozole, by Merck Group (1 mentions) Ruxolitinib, by Selleck Chemicals (1 mentions) Tamoxifen, by Merck Group (1 mentions) Disposable pd 10 desalting column, by GE Healthcare (1 mentions)

Close spelling variants of this product

MTT assay (11 citations) MTT cell viability assay (4 citations)

22 protocols using mtt cell viability assay kit

ACKR3 Silencing Optimization

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Lipofectamine 2000 (11668-019; Life Technologies, Invitrogen, Waltham, MA, USA) was used to inject ACKR3 siRNA (109229 and 112094; SilencerTM siRNA, Thermo Fisher Scientific) or negative control siRNA (4390843; Silencer™ Select Negative Control No. 1 siRNA, Thermo Fisher Scientific) into cells. RNA was collected from cells after 72 h, and specific gene knockdown was assessed by RT-qPCR. Cell viability after siRNA transfection was assessed using the MTT Cell Viability Assay Kit (Biotium, Inc., Fremont, CA, USA) as per the protocol recommended by the manufacturer.

Takaya K., Asou T, & Kishi K. (2022). Selective Elimination of Senescent Fibroblasts by Targeting the Cell Surface Protein ACKR3. International Journal of Molecular Sciences, 23(12), 6531.

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Macrophage Differentiation and Activation

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For NFI-A transfection, plasmid DNA was suspended in HiPerFect reagent at a 0.5 μg/ml final concentration (Qiagen, Valencia, CA, USA). For Rb knockdown, pools of Rb-specific or scrambled (control) siRNAs were suspended in HiPerFect reagent at a 0.5 μM final concentration. Cells were transfected using the Gene Pulser MXCell system (Bio-Rad, Hercules, CA, USA). After 24 h, cells were differentiated for 6 d with M-CSF plus rIL-4. In some experiments, differentiated cells were stimulated for 12 h with Gram-negative bacterial LPS (Escherichia coli serotype 0111:B4; Sigma-Aldrich, St. Louis, MO, USA). Cell viability was assessed by a MTT Cell Viability Assay Kit (Biotium, Fremont, CA, USA).

Dai J., Kumbhare A., Williams D.A., Youssef D., Yao Z.Q., McCall C.E, & Gazzar M.E. (2017). Nfia deletion in myeloid cells blocks expansion of myeloid-derived suppressor cells during sepsis. Innate Immunity, 24(1), 54-65.

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MTT Assay for Cell Viability

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The MTT cell viability assay kit (Biotium) was used to test metabolic function of cells. Following manufacturer’s recommendations, MDBK cells were seeded into a 96-well tissue culture plate (6 × 10³ cells per well). After completion of an experiment, cells were washed with PBS and 10 µL of MTT solution was added to the wells, mixed gently and incubated for 4 h at 37 °C in a 5% CO₂ atmosphere. 200 µL of dimethyl sulfoxide (DMSO) was added to the mixture to solubilize the tetrazolium salt that was produced by the metabolically active cells and absorbance was measured at 570 nm. Background absorbance was measured at 630 nm and subtracted from signal absorbance to yield normalized absorbance values.

Cowick C.A., Russ B.P., Bales A.R., Nanduri B, & Meyer F. (2022). Mannheimia haemolytica Negatively Affects Bovine Herpesvirus Type 1.1 Replication Capacity In Vitro. Microorganisms, 10(11), 2158.

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Macrophage Viability Assay

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THP-1-derived macrophages were seeded at 5×10⁴ cells/well in a 96-well plate. Cytotoxicity to SFN or S. aureus infection was assessed by an MTT cell viability assay kit (Biotium, Inc.) to measure cellular metabolic activity following the manufacturer's protocol. Absorbance changes were measured at 550 nm, and the background absorbance at 600 nm, using the FLUOstar Omega microplate reader (BMG Labtech).

Deramaudt T.B., Ali M., Vinit S, & Bonay M. (2020). Sulforaphane reduces intracellular survival of Staphylococcus aureus in macrophages through inhibition of JNK and p38 MAPK-induced inflammation. International Journal of Molecular Medicine, 45(6), 1927-1941.

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Propofol Exposure and Cell Viability

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To determine cell viability, BMECs were treated with 10, 50, 100, or 1,000 μM propofol in EC ± media at 37°C on a rotational platform. Following 3 h of treatment, propofol was aspirated and replaced with 100 μL of EC ± media. A MTT Cell Viability Assay Kit (Biotium, Fremont, CA, United States) was utilized to determine cell viability following propofol exposure. Following manufacturer’s instructions, 10 μL of MTT solution was added to each well and the plate was incubated at 37°C for 2 h on a rotational platform. After 2 h of incubation, 200 μL of DMSO (Sigma Aldrich, St. Louis, MO, United States) was added to each well, triturating several times to dissolve the formazan salt. Absorbance was measured at 570 nm using a Synergy HTX Multi-Mode reader and normalized by subtracting background absorbance measured at 630 nm.

Hughes J.M., Neese O.R., Bieber D.D., Lewis K.A., Ahmadi L.M., Parsons D.W, & Canfield S.G. (2022). The Effects of Propofol on a Human in vitro Blood-Brain Barrier Model. Frontiers in Cellular Neuroscience, 16, 835649.

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Cell Viability and Colony Formation Assays

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Cell viability was determined by MTT cell viability assay kit (Biotium Inc., Beijing, China). MCF-7 or MDA-MB-231 cells were seeded in 96-well plates and were incubated at 37 °C for 24 h to approximately 85% confluence, and then cells were treated with 0.1 mM lidocaine, 0.2 μM cisplatin or with both agents for various hours. Then cells were incubated with 10 μL MTT and were incubated at 37 °C for 4 h. Post addition of 200 μL DMSO into each well to dissolve the formazan, the absorbance was measured on an ELISA plate reader with a test wavelength of 570 nm and a reference wavelength of 630 nm to obtain sample signal (OD₅₇₀–OD₆₃₀). For cell colony formation assay, 4 × 10² cells were incubated in 6-well plates at 37 °C containing 5% CO₂, and were treated with 0.1 mM lidocaine, 0.2 μM cisplatin or with both agents. 96 h post treatment, cells were stained with crystal violet (0.005%) for 30 min and colony numbers were counted.

Li K., Yang J, & Han X. (2014). Lidocaine Sensitizes the Cytotoxicity of Cisplatin in Breast Cancer Cells via Up-Regulation of RARβ2 and RASSF1A Demethylation. International Journal of Molecular Sciences, 15(12), 23519-23536.

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Evaluating Antiproliferative Effects of Compounds

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NL and CLL cybrids were plated in a 96-well plate at a density of 10,000 cells per well in 100 μL of culture media. After 24 h plating, the cells were treated for 48 h with ibrutinib, ALA, amla, melatonin, and resveratrol at concentrations of 10 μM, 1 mM, 300 μg, 1 mM, and 100 μM, respectively. After the completion of the treatment, the cell metabolism was measured using the colorimetric MTT assay kit according to the manufacturer’s protocol (MTT Cell Viability Assay Kit, Biotium, Fremont, CA, USA). A minimal of three replicates were tested per sample. Each experiment was repeated three times.

Singh L., Atilano S., Chwa M., Singh M.K., Ozgul M., Nesburn A, & Kenney M.C. (2023). Using Human ‘Personalized’ Cybrids to Identify Drugs/Agents That Can Regulate Chronic Lymphoblastic Leukemia Mitochondrial Dysfunction. International Journal of Molecular Sciences, 24(13), 11025.

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MTT Assay for Cell Viability Assessment

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After the exposure time, cell metabolic activity was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay (Biotium, MTT Cell Viability Assay Kit, cat. no. 30006). The assay was performed according to the manufacturer’s instructions. Briefly, 10 µL of MTT agent was added to each well of the 96-well cell culture plate. The cell culture plates were then incubated in the dark for 4 h. After this period, 200 µL of dimethyl sulfoxide (DMSO, Corning Media Tech, Cat. no. 15303671) was added to each of the wells and resuspended to break up the crystals formed.
The absorbance was measured using a HEALES model MB-580 microplate reader (HEALES, Shenzhen, China) at wavelengths of 570 nm and 630 nm (corresponding to the background signal).
The deviation from the mean of the absorbance values obtained for each of the exposure configurations in each of the wells of the cell culture plate is calculated. The representation of the heat maps for the standard deviation values obtained with respect to the mean absorbance is performed with Prism 9 software (GraphPad^© Software, v.9.3.1., Massachusetts, United States).

Rivera González M.X., López de Mingo I., Amuneke Ramírez A, & Maestú Unturbe C. (2024). Design and characterisation of a cell exposure system with high magnetic field homogeneity: RILZ coils. Frontiers in Bioengineering and Biotechnology, 12, 1337899.

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Defensin Modulation of HIV Attachment and Entry

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Recombinant hBD2 and hBD3 were obtained from Santa Cruz Biotechnology, as well as from Dr. Aaron Weinberg’s laboratory. All experiments with hBD2 and hBD3 were performed in keratinocyte growth medium (Lonza). The toxicity of hBDs on polarized cells was examined by using an MTT Cell Viability Assay Kit (Biotium, Inc.). One set of cells was pretreated with antibodies against HSPG (25 mg/ml) or isotype control antibodies (25 µg/ml). Another set of cells was pretreated with 10 U/ml heparinase (Sigma) for 30 min. hBD2 and hBD3 were added to the apical surfaces of polarized cells, and cells were incubated at 4°C for 30 min. Cells were washed 3 times with PBS, pH 7.0, and dual (X4-R5)-tropic HIV-1_SF33, and R5-tropic HIV-1_SF170 or X4-tropic HIV-1_92UG029 virions (40 ng/ml of each) were added to the apical surfaces of the cells. For the HIV attachment assay, cells were incubated at 4°C for 1 h and then washed 3 times and lysed with Triton X-100 buffer. Surface-bound HIV was measured by ELISA. For HIV entry, cells were incubated with defensins at 4°C for 30 min and then washed and incubated with HIV at 37°C for 2 h. Uninternalized virions from the cell surface were removed with 0.25% trypsin as previously described (Tugizov et al., 2012 (link)). Cells were then lysed, and internalized virus was examined by using p24 ELISA (PerkinElmer).

Herrera R., Morris M., Rosbe K., Feng Z., Weinberg A, & Tugizov S. (2015). Human beta-defensins 2 and -3 cointernalize with human immunodeficiency virus via heparan sulfate proteoglycans and reduce infectivity of intracellular virions in tonsil epithelial cells. Virology, 487, 172-187.

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mADSCs Viability Assessment

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10⁴ mADSCs per well were seeded in MW96 and grown in the standard culture medium described above. Subsequently, they were treated or not treated with increasing concentrations (25 μM, 50 μM, 100 μM, 200 μM, 300 μM, 400 μM, 500 μM) of MGO for 16 h. The viability of the mADSCs was then assessed by using the “MTT Cell Viability Assay” kit (Biotium, Fremont, CA, USA) according to the manufacturer’s instructions. The sample was read using a spectrophotometer (Infinite 200, Tecan, Mannedorf, Switzerland) at 595 nm.

Leone A., Nicolò A., Prevenzano I., Zatterale F., Longo M., Desiderio A., Spinelli R., Campitelli M., Conza D., Raciti G.A., Beguinot F., Nigro C, & Miele C. (2023). Methylglyoxal Impairs the Pro-Angiogenic Ability of Mouse Adipose-Derived Stem Cells (mADSCs) via a Senescence-Associated Mechanism. Cells, 12(13), 1741.

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