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Celltiter glo 2.0 assay kit

Manufactured by Promega
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The CellTiter-Glo 2.0 assay kit is a luminescent cell viability assay that quantifies ATP, an indicator of metabolically active cells. The assay uses a proprietary thermostable luciferase to generate a stable luminescent signal proportional to the amount of ATP present in the sample.

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Spelling variants of this product

CellTiter-Glo (2548 citations) CellTiter-Glo assay (840 citations) CellTiter-Glo 2.0 (249 citations) CellTiter-Glo kit (240 citations) CellTiter-Glo 2.0 kit (43 citations)

69 protocols using celltiter glo 2.0 assay kit

1

Cellular ATP Level Quantification

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ATP level in cells was assessed using the luminometer (Promega, WI, USA) and CellTiter-Glo 2.0 Assay kits. The total protein in relative cell lysate was used to normalize the relative cellular ATP level [33 (link)].
Liu X., Zhao T., Yuan Z, & Ge S. (2022). MIR600HG sponges miR-125a-5p to regulate glycometabolism and cisplatin resistance of oral squamous cell carcinoma cells via mediating RNF44. Cell Death Discovery, 8, 216.
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2

Biomaterial Characterization and Cell Culture

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Branched polyethyleneimine (BPEI, MW=25,000), chitosan (CH, medium molecular weight), phosphate-buffered saline (PBS), paraformaldehyde, glutaraldehyde, menadione, filtered water for cell culture, fluorescein-labeled hyaluronic acid, chloramphenicol, and medical grade titanium disks were purchased from Sigma-Aldrich. Sodium hyaluronate (HA, MW 1,5000,000–2,200,000) was purchased from Acros Organic. 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT), RPMI 1640 powder containing L-glutamine and phenol red (without HEPES or Na bicarbonate), penicillin-streptomycin (10,000 U/mL), NaCl, 3-(N-morpholino)propanesulfonic acid (MOPS), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), N-hydroxysulfosuccinimide (Sulfo-NHS), Calcein-AM, ethidium homodimer-1, Hoechst 33342, Nunc™ Lab-Tek™ II Chamber Slide™ System, and Pierce™ Quantitative Fluorometric Peptide Assay were purchased from Thermo Fisher Scientific. α-MEM (1x) minus ascorbic acid was obtained from Gibco. Osmium tetroxide (4%) was obtained from Electron Microscopy Sciences. Accutase was purchased from Innovative Cell Technologies. Cell Titer Glo 2.0 assay kits were obtained from Promega. All materials were used as received.
Rodríguez López A.D., Lee M.R., Ortiz B.J., Gastfriend B.D., Whitehead R., Lynn D.M, & Palecek S.P. (2019). Preventing S. aureus biofilm formation on titanium surfaces by the release of antimicrobial β-peptides from polyelectrolyte multilayers. Acta biomaterialia, 93, 50-62.
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3

Measuring Cell Viability and Apoptosis

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Supernatants collected on day 3 after infection (or indicated otherwise) were analysed for HCG and LDH levels. HCG was measured using the Abnova HCG ELISA Kit (KA4005) according to the manufacturer’s instructions. All of the supernatants were diluted between 1/1,000 to 1/2,000 before analysis. LDH was measured using the Abcam LDH cytotoxicity kit II (ab65393) according to the manufacturer’s instructions in undiluted supernatants using an LDH standard curve. Cell viability was measured using the Promega Cell-Titer Glo 2.0 assay kit. Caspase 3/7 activity was measured using the Caspase Glo 3/7 Assay System (Promega) according to the manufacturer’s instructions.
Chen J., Neil J.A., Tan J.P., Rudraraju R., Mohenska M., Sun Y.B., Walters E., Bediaga N.G., Sun G., Zhou Y., Li Y., Drew D., Pymm P., Tham W.H., Rossello F.J., Nie G., Liu X., Subbarao K, & Polo J.M. (2023). A placental model of SARS-CoV-2 infection reveals ACE2-dependent susceptibility and differentiation impairment in syncytiotrophoblasts. Nature Cell Biology, 25(8), 1223-1234.
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4

Cell Viability Quantification via ATP Assay

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To determine cell viability, we used the CellTiter-Glo 2.0 assay kit (Promega) that quantifies the amount of ATP present, which indicates the percentage of metabolically active cells. Briefly, a cell culture plate was incubated at room temperature for 30 min. Next, 100 μl/well of the supernatant was removed, followed by the addition of 100 μl cell viability assay reagent. Samples were incubated for 10 min at 22°C. The luminescence level per well was determined as described above. Percent cell viability for each sample was calculated based on the percent luminescence relative to the DMEM control that contained 5% NBCS and 1% NEAA.
Özçam M., Tocmo R., Oh J.H., Afrazi A., Mezrich J.D., Roos S., Claesen J, & van Pijkeren J.P. (2019). Gut Symbionts Lactobacillus reuteri R2lc and 2010 Encode a Polyketide Synthase Cluster That Activates the Mammalian Aryl Hydrocarbon Receptor. Applied and Environmental Microbiology, 85(10), e01661-18.
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5

Dose-Dependent Cell Viability Assay

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Serially diluted aliquots of oHSV were prepared in a 96-well plate, and then single-cell suspension at the density of 20,000 cells per well was transferred to the new plates. After five-day incubation, the amount of ATPs was quantified using a CellTiter-Glo 2.0 Assay kit (Promega) and luminescent signals were read at 520 nm using a plate reader (POLARstar Omega; BMG Labtech). Data were normalized at maximum γ-axis values, followed by nonlinear regression analysis to plot the dose–response curves using Prizm 6 (GraphPad Software, Inc).
Nakashima H., Nguyen T., Kasai K., Passaro C., Ito H., Goins W.F., Shaikh I., Erdelyi R., Nishihara R., Nakano I., Reardon D.A., Anderson A.C., Kuchroo V, & Chiocca E.A. (2018). Toxicity and Efficacy of a Novel GADD34-expressing Oncolytic HSV-1 for the Treatment of Experimental Glioblastoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(11), 2574-2584.
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6

Measuring Intracellular ATP Levels

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The level of ATP in cells was measured using a luminometer and a CellTiter-Glo 2.0 Assay kit from Promega as per instructions. The total protein in the corresponding cell lysate was used to normalize the relative intracellular ATP level (measured as luminescence).
Zhang Z., Tan X., Luo J., Yao H., Si Z, & Tong J.S. (2020). The miR-30a-5p/CLCF1 axis regulates sorafenib resistance and aerobic glycolysis in hepatocellular carcinoma. Cell Death & Disease, 11(10), 902.
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7

Cell Proliferation Assay Protocol

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Cell proliferation was measured using the CellTiter-Glo 2.0 assay kit (G9243; Promega, Madison, WI, USA) and cell counting method. Cells were plated on a 96-well black plate for the CellTiter-Glo 2.0 assay, whereas a 24-well plate was used for cell counting. After seeding the cells for 24 h, they were treated with the indicated concentrations of PM, followed by either CellTiter-Glo 2.0 treatment or direct counting using a hemocytometer. The luminescence intensity of cells was measured using SpectraMax i3 (Molecular Devices, San Jose, CA, USA).
Jin S., Yoon S.J., Jung N.Y., Lee W.S., Jeong J., Park Y.J., Kim W., Oh D.B, & Seo J. (2023). Antioxidants prevent particulate matter-induced senescence of lung fibroblasts. Heliyon, 9(3), e14179.
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8

Cell Viability Assessment with MTT and CellTiter-Glo

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The cell survival assay was performed as previously described [50 (link)]. Briefly, cells were seeded in 96-well culture plates. After 24 h, cells were treated with drugs, and cell survival was measured by the MTT assay or by using the CellTiter-Glo® 2.0 Assay Kit (G7572, Promega) according to the manufacturer’s instructions. For the MTT assay, MTT reagent was added together with a different range of H2O2 for 4 h, the purple formazan crystal was dissolved with DMSO, and the colored solution was quantified using a plate reader. For the CellTiter-Glo® 2.0 Assay, cells were lysed with CellTiter-Glo® reagent and the luminescence signal was read using a plate reader.
Kim S., Kim Y., Kim Y., Yoon S., Lee K.Y., Lee Y., Kang S., Myung K, & Oh C.K. (2023). PCNA Ser46-Leu47 residues are crucial in preserving genomic integrity. PLOS ONE, 18(5), e0285337.
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9

Neutralization of EV71 Clinical Strain

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Neutralizing activity of plant-produced cD5 antibody against EV71 clinical strain EV71/G082 was measured using the micro-neutralization assay as described previously [44 (link)]. After incubation for three days, cell viability was measured using the CellTiter-Glo 2.0 assay kit (Promega, Fitchburg, WI, USA) following the manufacturer’s protocol. The percentage of neutralization was calculated as follows: 100 × (luminescence of the given sample—luminescence of the virus-only sample)/(luminescence of the cell-only sample—luminescence of the virus-only sample). Half-maximal inhibitory concentration (IC50) of plant-produced cD5 antibody was determined using variable-slope nonlinear regression analysis by GraphPad Prism software.
Rattanapisit K., Chao Z., Siriwattananon K., Huang Z, & Phoolcharoen W. (2019). Plant-Produced Anti-Enterovirus 71 (EV71) Monoclonal Antibody Efficiently Protects Mice Against EV71 Infection. Plants, 8(12), 560.
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10

Cell Viability Assays with H2O2

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Cells were seeded in 96-well culture plates. After 24 h, cells were treated with drugs and cell survival was measured by the MTT assay or by using the CellTiter-Glo® 2.0 Assay kit (Promega) according to the manufacturer’s instructions. For the MTT assay, MTT reagent was added together with a different range of H2O2 for 4 h, the purple formazan crystal was dissolved with DMSO, and the colored solution was quantified using a plate reader. For the CellTiter-Glo® 2.0 Assay, in brief, cells were lysed with CellTiter-Glo® reagent and the luminescence signal was read using a plate reader. Percent cell survival was normalized to that of control cells.
Park S.H., Kim Y., Ra J.S., Wie M.W., Kang M.S., Kang S., Myung K, & Lee K.Y. (2021). Timely termination of repair DNA synthesis by ATAD5 is important in oxidative DNA damage-induced single-strand break repair. Nucleic Acids Research, 49(20), 11746-11764.
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