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Celltitre glo luminescent assay

Manufactured by Promega
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CellTiter-Glo® Luminescent Assay is a cell viability assay that quantifies the amount of ATP present in metabolically active cells. The assay provides a homogeneous, luminescent method for determining the number of viable cells in culture.

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Lab products found in correlation

Mek162, by Selleck Chemicals (1 mentions) Bay11 7082, by Merck Group (1 mentions) Sp600125, by Merck Group (1 mentions) Prism 6, by GraphPad (1 mentions) Ku55933, by Selleck Chemicals (1 mentions) Nu7441, by Selleck Chemicals (1 mentions) 5 ethynyl 2 deoxyuridine (edu), by Thermo Fisher Scientific (1 mentions) Envision plate reader, by PerkinElmer (1 mentions)

Close spelling variants of this product

CellTitre-Glo® Luminescent Cell Viability Assay kit CellTitre-Glo® 3-D luminescent assay CellTitre-Glo luminescent cell viability assay CellTitre-Glo® Luminescent 3D Cell Viability Assay CellTitre-Glo assay CellTitre-Glo Luminescent assays

3 protocols using celltitre glo luminescent assay

1

Cytotoxicity Assay with Inhibitors

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Cells were seeded in complete growth medium at a density of 2000–4000 cells/well in 96-well tissue culture plates based on their cell doubling times. Cells were pre-treated with Jun kinase inhibitor (JNKi) SP600125, NFκB inhibitor (BAYi) BAY-11-7082 (Sigma) or MEK1/2 inhibitor (MEKi) MEK162 (Selleckchem, Houston, TX, USA) for 24 h. After serial dilutions, 100 μl of complete growth medium containing 5-Fluorouracil, cisplatin, paclitaxel or doxorubicin were added to cells in increasing amounts. DMSO was used as controls. Plates were incubated for 72 h after which cell viability was assessed by CellTitre-Glo® Luminescent Assay according to the manufacturer’s instructions (Promega, Madison, WI, USA). Reactions were carried out in triplicate and analyzed with GraphPad Prism 6 (GraphPad Software Inc., San Diego, CA, USA).
Rasheed S.A., Leong H.S., Lakshmanan M., Raju A., Dadlani D., Chong F.T., Shannon N.B., Rajarethinam R., Skanthakumar T., Tan E.Y., Hwang J.S., Lim K.H., Tan D.S., Ceppi P., Wang M., Tergaonkar V., Casey P.J, & Iyer N.G. (2017). GNA13 expression promotes drug resistance and tumor-initiating phenotypes in squamous cell cancers. Oncogene, 37(10), 1340-1353.
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2

Quantifying DNA Replication and ATP Levels

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Chemical inhibitors KU55933 and NU7441 (Selleckchem) were dissolved in DMSO and stored as 2 mM stock. Each inhibitor (10 or 20 μM), dissolved in F12-K media, was incubated with A549 cells for up to 6 h. To detect DNA replication, 10 μM EdU (Invitrogen) was incubated in F12-K medium for 30 min at 37 °C. ATP level in cell culture was determined in 96-well plate by using Cell Titre-Glo luminescent assay (Promega).
Rai P., He F., Kwang J., Engelward B.P, & Chow V.T. (2016). Pneumococcal Pneumolysin Induces DNA Damage and Cell Cycle Arrest. Scientific Reports, 6, 22972.
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3

Quantifying Cytotoxicity Using CellTitreGlo Assay

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The CellTitreGlo® luminescent assay (Promega, Madison Wisconsin, USA) was used to assess cytotoxicity by quantifying the loss of intracellular ATP relative to the vehicle controls. LUHMES, SH-SY5Y or NSC cells were grown in 384 well plates to 1 × 104 cells/well and subjected to differentiation for seven days. Cells were treated for 24 hours, then 30 μL CellTitreGlo® luminescent substrate was added and incubated for 30–60 minutes at room temperature. Luminescence was read in an EnVision plate reader (Perkin Elmer, Waltham MA, USA). 0.5% DMSO treated control wells were used to determine 100% viability, and data from each experiment were normalized to the mean of these control wells. Concentration-response was plotted for each toxicant:cell-type combination, and the IC50 was identified by interpolation as the concentration that reduced intracellular [ATP] to 50% of control. CellTitreGlo® assays were carried out in quadruplicate for Figures 1, 2A–C, 3, 4, and 6, and all experiments were repeated to confirm the reproducibility of results.
Tong Z.B., Kim H., El Touny L., Simeonov A, & Gerhold D. (2022). LUHMES Dopaminergic Neurons are Uniquely Susceptible to Ferroptosis. Neurotoxicity research, 40(5), 1526-1536.
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