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Celltiter 96 aqueous solution cell proliferation assay kit

Manufactured by Promega
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The CellTiter 96 Aqueous Solution Cell Proliferation Assay kit is a colorimetric method for determining the number of viable cells in a proliferation or cytotoxicity assay. The assay uses a tetrazolium compound [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS] and an electron coupling reagent (phenazine methosulfate; PMS) to produce a colored formazan product that is soluble in tissue culture medium.

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Lab products found in correlation

Cisplatin, by Merck Group (1 mentions) Tlc plate, by Merck Group (1 mentions) Sephadex lh 20 resin, by Merck Group (1 mentions) Spectramax 190, by Molecular Devices (1 mentions) Mueller hinton medium, by BD (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Rpmi 1640 medium, by Merck Group (1 mentions) Dimethyl sulfoxide (dmso), by Avantor (1 mentions)

Close spelling variants of this product

CellTiter 96 AQueous CellTiter 96 kit CellTiter 96

6 protocols using celltiter 96 aqueous solution cell proliferation assay kit

1

MTS-based Cell Viability Assay

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Cell viability was assayed by a Cell Titer 96 Aqueous Solution Cell Proliferation Assay Kit (Promega, Madison, WI, USA). Myoblasts were seeded at a density of 5 × 103 (200 μL per well) in 96-well plates and cultured as described above, with five replicates for each group of samples (n = 5). Then, 20 μL of MTS (3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)2H-tetrazolium) was added to each well, and the cells were incubated in the dark at 37 °C for 3 h followed by measuring the absorbance of formazan at a wavelength of 490 nm on a multimode microplate reader (Biotech, Winooski, VT, USA).
Zhang S., Qiu X., Zhang Y., Huang C, & Lin D. (2023). Metabolomic Analysis of Trehalose Alleviating Oxidative Stress in Myoblasts. International Journal of Molecular Sciences, 24(17), 13346.
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2

Cisplatin Cytotoxicity Assay in 96-well Plates

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Cells cultured in 96-well plates (3,000 cells/well) were incubated with cisplatin (Sigma-Aldrich, USA) diluted directly in the culture medium in concentrations varying from 2.5 to 40 µM. After 24 and 48 h, cell viability was determined with MTS assay (CellTiter 96® Aqueous Solution Cell Proliferation Assay kit, Promega, USA) according to manufacturer’s protocol.
Dourado M.R., Elseragy A., da Costa B.C., Téo F.H., Guimarães G.N., Machado R.A., Risteli M., Wahbi W., Gurgel Rocha C.A., Paranaíba L.M., González-Arriagada W.A., da Silva S.D., Rangel A.L., Marques M.R., Rossa Junior C., Salo T, & Coletta R.D. (2023). Stress induced phosphoprotein 1 overexpression controls proliferation, migration and invasion and is associated with poor survival in oral squamous cell carcinoma. Frontiers in Oncology, 12, 1085917.
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3

Cell Viability and Colony Formation

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Cell viability was determined at 24, 48 and 72 hours using a CellTiter 96 Aqueous Solution Cell Proliferation Assay kit (Promega, Madison, WI) according to the manufacturer’s instructions. After transfection for 72 hours, cells were seeded at a low density (1000 cells/plate) for colony formation assay and were allowed to grow until visible colonies were formed. Plates were then stained with giemsa and colonies were counted.
Dasgupta P., Kulkarni P., Majid S., Varahram S., Hashimoto Y., Bhat N.S., Shiina M., Deng G., Saini S., Tabatabai Z.L., Yamamura S., Tanaka Y, & Dahiya R. (2018). MicroRNA-203 inhibits long noncoding RNA HOTAIR and regulates tumorigenesis through epithelial-to-mesenchymal transition pathway in renal cell carcinoma. Molecular cancer therapeutics, 17(5), 1061-1069.
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4

Purification and Bioactivity of Coriolic Acid

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Silica gel 60A (Analtech, Newark, DE, USA) and silica thin-layer chromatography (TLC) plates were purchased from Merck KGaA (Darmstadt, Germany), and Sephadex LH-20 resin was purchased from Sigma (St. Louis, MO, USA). High-performance liquid chromatography (HPLC) was performed on a Shimadzu 20A system (Shimadzu, Kyoto, Japan). Cell growth was assayed using the CellTiter 96® Aqueous Solution cell proliferation assay kit (Promega, WI, USA). We obtained coriolic acid by purification of S. herbacea L. extracts.
Ko Y.C., Choi H.S., Kim J.H., Kim S.L., Yun B.S, & Lee D.S. (2020). Coriolic Acid (13-(S)-Hydroxy-9Z, 11E-octadecadienoic Acid) from Glasswort (Salicornia herbacea L.) Suppresses Breast Cancer Stem Cell through the Regulation of c-Myc. Molecules, 25(21), 4950.
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5

Colorimetric Assay for Cell Viability and Clonogenicity

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Cell viability of established stable and their respective control cells as well as cells transiently transfected with siRNAs and miR-155 mimic was determined at 24, 48, and 72 hours using a CellTiter 96 Aqueous Solution Cell Proliferation Assay kit (Promega, Madison, WI) according to the manufacturer’s instructions. This is a colorimetric assay which measures the activity of reductase enzymes. Briefly cells were seeded at a density of 1.2×103 for Caki-1 and 1×103 for 786-O per well in flat bottomed 96-well plates. At indicated times, CellTiter 96 Aqueous One reagent was added to each well and absorbance was measured at 490nm using SpectraMAX 190 (Molecular Device, Sunnyvale, CA).
For colony-forming assays, stable Caki-1 and 796-O anti-miR-155 / TCL6-OE cells and their respective control cells were plated at a low density onto 10cm dishes (1000 cells/plate). These cells were allowed to grow until visible colonies were formed.
Similarly for cells with transient miR-155 overexpression and suppression of lncTCL6, 72 hrs post-transfection, cells were counted and seeded at low density. Following 8 days (786-O) or 12 days (Caki-1) of cell adherence, colonies of cells were fixed with Geimsa stain and further stained with crystal violet reagent. The colonies were counted and recorded.
Kulkarni P., Dasgupta P., Hashimoto Y., Shiina M., Shahryari V., Tabatabai Z.L., Yamamura S., Tanaka Y., Saini S., Dahiya R, & Majid S. (2021). A lncRNA TCL6-miRNA-155 interaction regulates the Src-Akt-EMT network to mediate kidney cancer progression and metastasis. Cancer research, 81(6), 1500-1512.
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6

Antibacterial and Cytotoxic Activity Assays

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For antibacterial activity, double-distilled water, dimethyl sulfoxide (DMSO) (JT Baker, USA), INC-80 incubator (Prendo), sterile 96-well round-bottom microplates with lid (Corning Costar, New York), and Mueller Hinton medium were used (Becton Dickinson). Strains of drug-resistant clinical isolates were obtained from the University Hospital Dr. Eleuterio González of the Universidad Autónoma de Nuevo León. Four of these bacteria are included in the list of priority pathogens issued by the WHO, 2017 . For cytotoxic activity, the cell lines tested were obtained from ATCC (American Type Culture Collection, Manassas, VA, USA), RPMI-1640 medium (Sigma Aldrich, St. Louis, MO, USA), DMEM (Invitrogen, Thermo Fisher Scientific, Inc., Waltham, MA, USA) and fetal bovine serum (SFB, Invitrogen), Cell Titer 96® aqueous solution cell proliferation assay kit (Promega, Madison, WI, USA), automated ELISA reader.
Nocedo-Mena D., Rivas-Galindo V.M., Navarro P., Garza-González E., González-Maya L., Ríos M.Y., García A., Ávalos-Alanís F.G., Rodríguez-Rodríguez J, & Camacho-Corona M.D. (2020). Antibacterial and cytotoxic activities of new sphingolipids and other constituents isolated from Cissus incisa leaves. Heliyon, 6(8), e04671.
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