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Celltiter 96 aqueous one solution kit

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The CellTiter 96 AQueous One Solution kit is a colorimetric assay for determining the number of viable cells in a cell proliferation or cytotoxicity assay. The kit contains a tetrazolium compound that is bioreduced by cells into a colored formazan product, which can be quantified using a spectrophotometer.

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Lab products found in correlation

Synergy ht, by Agilent Technologies (3 mentions) Microplate reader, by Bio-Rad (3 mentions) Aldefluor kit, by STEMCELL (3 mentions) Model 680, by Bio-Rad (2 mentions) Leptin, by R&D Systems (2 mentions) Versamax, by Molecular Devices (2 mentions) Thp 1, by Korean Cell Line Bank (2 mentions) 96 well flat bottomed plate, by Corning (2 mentions) Live dead viability cytotoxicity kit, by Thermo Fisher Scientific (2 mentions) Celltiter 96 aqueous one solution, by Agilent Technologies (1 mentions) Leptin, by Merck Group (1 mentions) Medium 199, by Thermo Fisher Scientific (1 mentions) Huvecs, by Lonza (1 mentions) Hela cell, by American Type Culture Collection (1 mentions) Glomax multi multiplate reader, by Promega (1 mentions) A1 confocal microscope, by Nikon (1 mentions) Synergy h4 microplate reader, by Agilent Technologies (1 mentions) Prism 8, by GraphPad (1 mentions) Ix81 light microscope, by Olympus (1 mentions) Nf κb p65 transcription factor assay kit, by Abcam (1 mentions)

44 protocols using celltiter 96 aqueous one solution kit

1

Cytotoxicity Evaluation of Chloroquine

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Cytotoxicity measurements were based on the viability of Vero E6 cells in the presence of various concentrations of chloroquine as previously described (Keyaerts et al., 2004 (link)). After a 5-day incubation period, the number of viable cells was quantified by a tetrazolium-based colorimetric method, in which the reduction of MTS (CellTiter 96 AQueous One Solution kit, Promega, Netherlands) by mitochondrial dehydrogenases to a soluble colored formazan was measured in a spectrophotometer at 492 nm. The cytotoxic concentration was defined as the concentration of the compound that reduced cell viability by 50% (50% cytotoxic concentration, CC50).
Vergote V., Laenen L., Mols R., Augustijns P., Van Ranst M, & Maes P. (2021). Chloroquine, an Anti-Malaria Drug as Effective Prevention for Hantavirus Infections. Frontiers in Cellular and Infection Microbiology, 11, 580532.
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2

MTS Assay for CuB-induced Proliferation

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Cell proliferation was measured by MTS assay using the CellTiter 96 Aqueous ONE Solution kit (Promega, Madison, WI). Briefly, cells were seeded into 96-well plates at a density of 4×104/ml and 100 μl/well for 24 h. On the next day, culture medium was replaced. A375 cells were treated with CuB (0.01 μM, 0.1 μM and 1 μM) alone or pretreated with SP600125 (20 μM) for 1 h followed by CuB treatment for 48 h then measured using MTS assays. MTS reagent (20 μl) was added to each well and incubated at 37°C for 1 h–4 h. The absorbance at 490 nm was measured using a microplate reader (Model 680; Bio-Rad, Richmond, CA). Three independent experiments were performed, each in triplicates.
Zhang Y.T., Xu L.H., Lu Q., Liu K.P., Liu P.Y., Ji F., Liu X.M., Ouyang D.Y, & He X.H. (2014). VASP Activation via the Gα13/RhoA/PKA Pathway Mediates Cucurbitacin-B-Induced Actin Aggregation and Cofilin-Actin Rod Formation. PLoS ONE, 9(4), e93547.
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3

Cell Viability Assay Using MTS

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MTS assay was performed according to the standard protocol25 (link). The cultured cells were resuspended and seeded into 96-well plates (8x103 cells/well) and were incubated at different time points. Cell viability was analyzed with MTS using the Cell Titer 96® Aqueous One Solution kit (Promega Corporation) as determined by the manufacturer's protocol.
Han S., Chen S., Wang J., Huang S., Xiao Y, & Deng G. (2024). Erianin promotes apoptosis and inhibits Akt-mediated aerobic glycolysis of cancer cells. Journal of Cancer, 15(8), 2380-2390.
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4

Pomegranate Extract Cytotoxicity Assay

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Cytotoxicity was quantified with the CellTiter 96® AQueous One Solution kit (Promega Corp., Madison, WI, USA), assessing cell metabolic fitness through MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt]. Briefly, cells (5x103) were seeded in 96-well plates and left adhering for 24 h (at 37°C). Then, cells were treated for additional 24 h (in serum-free media) with 100 μg/mL of the indicated pomegranate extracts or the vehicle control (DMSO). At the end of treatments, 20 μL/well of the CellTiter 96® AQueous One Solution Reagent was added, cultures incubated for 1 h (at 37°C) and the absorbance (at 492 nm) was measured with a spectrophotometer (Synergy HT, BioTek).
Russo V., Continella A., Drago C., Gentile A., La Malfa S., Leotta C.G., Pulvirenti L., Ruberto G., Pitari G.M, & Siracusa L. (2021). Secondary metabolic profiles and anticancer actions from fruit extracts of immature pomegranates. PLoS ONE, 16(8), e0255831.
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5

Leptin-Induced Cell Proliferation Assay

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The cell proliferation assay was performed using the CellTiter 96 AQueous One Solution kit (Promega, Madison, WI, USA). In brief, cells were seeded at a concentration of 4 × 103 cells per well in 96-well plates. After cell attachment, the medium was replaced with DMEM containing 2% fetal bovine serum. After incubated in 2% fetal bovine serum DMEM for 48 h, cells were exposed to variable concentrations (1–100 ng ml−1) of leptin (R&D Systems, Inc., Minneapolis, MN, USA) for another 72 h, respectively. Cells were treated with leptin alone or/and pretreated with FAEE (Sigma–Aldrich, St Louis, MO, USA) for 1 h. Relative cell numbers were investigated by incubating cells with 20 μl CellTiter 96 AQueous One Solution Reagent for 4 h at 37 °C and then record the absorbance at 490 nm using a 96-well plate reader.
Tsai Y.C., Lee Y.M., Hsu C.H., Leu S.Y., Chiang H.Y., Yen M.H, & Cheng P.Y. (2015). The effect of ferulic acid ethyl ester on leptin-induced proliferation and migration of aortic smooth muscle cells. Experimental & Molecular Medicine, 47(8), e180-.
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6

Antimicrobial and Cytotoxicity Assays

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MICs were performed in 96-well microtiter plates. A DMSO stock solution of each compound was added to the first column and serially diluted across the columns of the plate. The last column of the plate contained no drug and served as a no-drug control. Overnight cultures of the bacterium being tested (MRSA ATCC 43300, MSSA ATCC 25923, or the aforementioned VRSA/VISA strains) were diluted 1000 fold (2 x 103 cells) and 100 μL was used as an inoculum in each well. MICs were determined by visual inspection for a pellet after 18 h incubation at 37 °C. Compounds were tested for cytotoxicity against Vero cells using the CellTiter 96 AQueous One Solution kit (Promega). Vero cells were seeded in 96 well plates at a density of 2x104 cells per well and the plates were incubated for 4 h at 37 °C to allow attachments of the Vero cells. Compounds were then added to the wells starting from a final concentration of 50 μg/mL and making twelve 1:2 dilutions. Cells were incubated for 72 h at 37 °C. Then 20 μL of freshly prepared MTS:PMS reagents were added to each well. The plates were incubated for 2 h and then read at an absorbance of 490 nm.
Patel J.S., Norambuena J., Al-Tameemi H., Ahn Y.M., Perryman A.L., Wang X., Daher S.S., Occi J., Russo R., Park S., Zimmerman M., Ho H.P., Perlin D.S., Dartois V., Ekins S., Kumar P., Connell N., Boyd J.M, & Freundlich J.S. (2021). Bayesian Modeling and Intrabacterial Drug Metabolism Applied to Drug-resistant Staphylococcus aureus. ACS infectious diseases, 7(8), 2508-2521.
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7

Evaluating HUVEC and Cardiomyocyte Viability

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Human umbilical vascular endothelial cells (HUVECs; Lonza, Basel, Switzerland) were seeded onto a collagen-coated plate and incubated in medium 199 (Invitrogen) supplemented with 20% FCS for 24 h. Neonatal rat cardiomyocytes were isolated from Lewis rats on postnatal day 1, as described previously [11] (link), and seeded onto a laminin-coated plate followed by incubation in α-MEM supplemented with 10% FCS for 24 h. Cells were then subjected to serum deprivation with/without hypoxia (1% O2) by culturing with serum-free medium or serum-free conditioned medium obtained from FM-MSCs cultured for 24 h. The cellular level of 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazo-lium (MTS), indicative of cell viability, as well as caspase-3 activity, was measured with a CellTiter96 AQueous One Solution Kit (Promega, Madison, WI) and a CaspACE™ Assay System Kit (Promega), according to the manufacturer’s instructions.
Yamahara K., Harada K., Ohshima M., Ishikane S., Ohnishi S., Tsuda H., Otani K., Taguchi A., Soma T., Ogawa H., Katsuragi S., Yoshimatsu J., Harada-Shiba M., Kangawa K, & Ikeda T. (2014). Comparison of Angiogenic, Cytoprotective, and Immunosuppressive Properties of Human Amnion- and Chorion-Derived Mesenchymal Stem Cells. PLoS ONE, 9(2), e88319.
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8

HeLa Cell Viability with Hydrogel Particles

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HeLa cell (ATCC, Manassas, VA, USA) viability in the presence and absence of hydrogel particles was measured by the methyl tetrazolium salt (MTS) assay. HeLa cells were seeded at a density of 5×103 per well (100 μL) in flat-bottomed 96-well plates (Corning Costar, Cambridge, MA, USA). The cells were next treated with 0.05, 0.1, 0.5, or 1 μg/mL hydrogel particles for 4 or 24 h. Untreated cells were used as controls. For the MTS assay, the CellTiter 96 AQueous One Solution kit (Promega, Madison, WI, USA) was used, following the manufacturer’s protocols. Briefly, the MTS reagent was added (10 μL per well), and the plates were incubated for 4 h at 37°C. The absorbance was detected at 490 nm with a microplate reader (VersaMax™; Molecular Devices, Sunnyvale, CA, USA). All of the experiments were repeated three times independently.
Cho S.H., Hong J.H., Noh Y.W., Lee E., Lee C.S, & Lim Y.T. (2016). Raspberry-like poly(γ-glutamic acid) hydrogel particles for pH-dependent cell membrane passage and controlled cytosolic delivery of antitumor drugs. International Journal of Nanomedicine, 11, 5621-5632.
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9

PLGA NPs Cytotoxicity Evaluation

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BMDCs (5×104 cells/100 μL per well) were treated with PLGA and PEI-coated PLGA NPs for 24 hours in flat-bottomed 96-well plates (Corning Incorporated, Corning, NY, USA). For the methyl tetrazolium salt (MTS) assay, the CellTiter 96 aqueous one-solution kit (Promega Corporation, Fitchburg, WI, USA) was used, following the manufacturer’s protocol. Briefly, the MTS reagent was added (10 μL per well), and the plates were incubated at 37°C for 3 hours. Absorbance was determined at 490 nm on a microplate reader (VersaMax; Molecular Devices, Sunnyvale, CA, USA).
Song C., Noh Y.W, & Lim Y.T. (2016). Polymer nanoparticles for cross-presentation of exogenous antigens and enhanced cytotoxic T-lymphocyte immune response. International Journal of Nanomedicine, 11, 3753-3764.
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10

MTS Cell Proliferation Assay in Monolayer

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MTS cell proliferation assay in monolayer was performed using CellTiter 96® AQueous One Solution kit (Promega, G3582), following the manufacturer’s protocol.
Villasante A., Robinson S.T., Cohen A.R., Lock R., Guo X.E, & Vunjak-Novakovic G. (2021). Human Serum Enhances Biomimicry of Engineered Tissue Models of Bone and Cancer. Frontiers in Bioengineering and Biotechnology, 9, 658472.
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