Celltiter glo luminescence assay

Manufactured by Promega

Sourced in United States

The CellTiter-Glo luminescence assay is a reagent-based kit designed to quantify the amount of ATP present in a sample, which is an indicator of metabolically active cells. The assay uses a luciferase reaction to generate a luminescent signal proportional to the amount of ATP in the sample.

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Lab products found in correlation

Prism 5, by GraphPad (5 mentions) Victor3 multilabel reader, by PerkinElmer (3 mentions) 96 well ultralow attachment plates, by Greiner (2 mentions) 96 well micro assay plate, by Greiner (2 mentions) Prism 6, by GraphPad (2 mentions) Spectramax m3, by Molecular Devices (2 mentions) Prism 9, by GraphPad (2 mentions) Accumax cell dissociation solution, by Innovative Cell Technologies (2 mentions) Infinite 200 pro instrument, by Tecan (2 mentions) Intesticult, by STEMCELL (2 mentions) Caspase glo 3 7 assay, by Promega (2 mentions) Envision plate reader, by PerkinElmer (2 mentions) Clariostar reader, by BMG Labtech (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Cytotox red, by Sartorius (1 mentions) Annexin 5, by Abcam (1 mentions) Prism software, by GraphPad (1 mentions) Spectramax m2e, by Molecular Devices (1 mentions) Facsdiva, by BD (1 mentions) Lsrfortessa, by BD (1 mentions)

Spelling variants of this product

CellTiter-Glo (2548 citations) CellTiter-Glo assay (840 citations) CellTiter-Glo luminescence (13 citations)

77 protocols using celltiter glo luminescence assay

ALDH1-high Cell Viability Assay

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Tumor-spheres were cultured as previously described [39 (link)]. ALDH1^high cells (1 × 10³/well) were cultured in ultralow attachment 96-well plates (Greiner) and treated with TLSC702 for 6 days. CellTiter-Glo^® luminescence assays (Promega) were performed with a TR717 Micro plate Luminometer (TROPIX) using a 96-well Micro-assay-plate (Greiner). Numerical values for the test groups are expressed to the 0.6% DMSO-treated group.

Tamori S., Nozaki Y., Motomura H., Nakane H., Katayama R., Onaga C., Kikuchi E., Shimada N., Suzuki Y., Noike M., Hara Y., Sato K., Sato T., Yamamoto K., Hanawa T., Imai M., Abe R., Yoshimori A., Takasawa R., Tanuma S.I, & Akimoto K. (2018). Glyoxalase 1 gene is highly expressed in basal-like human breast cancers and contributes to survival of ALDH1-positive breast cancer stem cells. Oncotarget, 9(92), 36515-36529.

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Cell Viability Assessment Protocols

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To assess cell viability, 10,000 (22Rv1), 20,000 cells (DuCaP/VCaP), or 500 cells (MIA-PaCa-2) were cultured in 96-well culture plates. Transfection with oligonucleotides was done 24 h after seeding. Four days after transfection, cell viability was measured using 3-(4,5-dimethylthiazol-2-yl)−2,5-dephenyltetrazolium bromide (MTT, 1 mg/ml) assays. Alternatively, cell viability was measured using CellTiter-Glo luminescence assays (Promega), following the manufacturer’s instructions. Absorbance (at 490 nm) and luminescence were measured using a Victor3 multilabel reader (PerkinElmer). Medium only was used as background control. To calculate the relative cell viability, cell viability values for each condition were normalized to the average of the cell viability values for control oligo-transfected cells. Each experiment was performed in triplicate and repeated at least three times.

Luna Velez M.V., Verhaegh G.W., Smit F., Sedelaar J.P, & Schalken J.A. (2019). Suppression of prostate tumor cell survival by antisense oligonucleotide-mediated inhibition of AR-V7 mRNA synthesis. Oncogene, 38(19), 3696-3709.

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Tumor Sphere Formation and Viability Assay

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Tumor-spheres were grown as previously described (21 (link),41 (link),42 (link),53 (link)). Briefly, cells (1×10³/well) were cultured in 96-well ultralow attachment plates (Greiner Bio-One) and treated with inhibitors for 6 days. Images were taken through an inverted microscope (Leica Microsystems, Inc.), and the numbers of tumor-spheres ≥50 µm in diameter were counted. Numerical values of the test groups are shown relative to the untreated cells. Cell Titer-Glo^® luminescence assays (Promega Corporation) were performed according to the manufacturer's protocol, and as previously described (21 (link),53 (link)). Values for test groups are shown relative to cells in the absence of the drug.

Motomura H., Ozaki A., Tamori S., Onaga C., Nozaki Y., Waki Y., Takasawa R., Yoshizawa K., Mano Y., Sato T., Sasaki K., Ishiguro H., Miyagi Y., Nagashima Y., Yamamoto K., Sato K., Hanawa T., Tanuma S.I., Ohno S, & Akimoto K. (2021). Glyoxalase 1 and protein kinase Cλ as potential therapeutic targets for late-stage breast cancer. Oncology Letters, 22(1), 547.

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Determining Cell Viability and Apoptosis

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For CellTiter-Glo luminescence assays (Promega), cells in 96-well plates were manually dosed or dosed using a D3000e Digital dispenser (HP Life Science Dispensing), and cell viability was determined 72–96 hours later. Percentage growth was determined using the equation (T – T0)/(C – T0) × 100, where T = compound-treaded cells; T0 = cells at 0-hour time point, and C = control cells. Incucyte cell confluence, annexin V green apoptosis, and Cytotox red (Essen BioScience, now Sartorius) assays were performed according to the manufacturer’s instructions. Cell growth and relative apoptosis/cell death were calculated as previously described (63 (link)).

Winkler C., King M., Berthe J., Ferraioli D., Garuti A., Grillo F., Rodriguez-Canales J., Ferrando L., Chopin N., Ray-Coquard I., Delpuech O., Bedognetti D., Ballestrero A., Leo E, & Zoppoli G. (2021). SLFN11 captures cancer-immunity interactions associated with platinum sensitivity in high-grade serous ovarian cancer. JCI Insight, 6(18), e146098.

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Flow Cytometric Analysis of Cell Death

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H1975, HCC827, PC9, ECLC26 cells were plated in 12-well plates at 8 × 10⁴ or 10⁵ cells / well and treated with the indicated agents next day. At the indicated time points, cell death was quantified by Annexin V (Bio Vision) staining, followed by flow cytometric analyses using a LSRFortessa (BD Biosciences). Data were analyzed using FACSDiva (BD biosciences). For EC₅₀ determination, cell viability was assessed by the CellTiter-Glo luminescence assays (Promega) using 96-well plates and a luminescent plate reader (SpectraMax M2e, Molecular Devices). EC50 value was calculated using Prism software (GraphPad).

Tanaka K., Yu H.A., Yang S., Han S., Selcuklu S.D., Kim K., Ramani S., Ganesan Y.T., Moyer A., Sinha S., Xie Y., Ishizawa K., Osmanbeyoglu H.U., Lyu Y., Roper N., Guha U., Rudin C.M., Kris M.G., Hsieh J.J, & Cheng E.H. (2021). Targeting Aurora B Kinase Prevents and Overcomes Resistance to EGFR Inhibitors in Lung Cancer by Enhancing BIM- and PUMA-mediated Apoptosis. Cancer cell, 39(9), 1245-1261.e6.

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Assessing Cell Proliferation with CellTiter-Glo®

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Cell proliferation was measured using the CellTiter-Glo^® luminescence assay (Promega, Madison, WI, USA) [29 (link)]. Huh7-derived LCSCs were added to 96-white well culture plates at 3 × 10³ cells per well and exposed to different concentrations (0–500 µg/mL) of HDT extracts. Following incubation for 7 days, 20 µL of substrate solution was added to each well, and luminescence was measured using a microplate reader (BioTek, Inc., Winooski, VT, USA). IC₅₀ values were analyzed with the curve-fitting program in GraphPad Prism 6 (GraphPad Software, La Jolla, CA, USA).

Kwon M, & Jung H.J. (2023). Hovenia dulcis Suppresses the Growth of Huh7-Derived Liver Cancer Stem Cells by Inducing Necroptosis and Apoptosis and Blocking c-MET Signaling. Cells, 13(1), 22.

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ATP Quantification in Permeabilized Cells

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Permeabilized and live cells in TRB were lysed and ATP levels quantified using the CellTiter-Glo luminescence assay (Promega) according to the manufacturers’ instructions. Luminescence was measured using a SpectraMax M3 microplate reader (Molecular Devices).

Duan L., Zaepfel B.L., Aksenova V., Dasso M., Rothstein J.D., Kalab P, & Hayes L.R. (2022). Nuclear RNA binding regulates TDP-43 nuclear localization and passive nuclear export. Cell reports, 40(3), 111106.

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Cellular Viability Assay Protocol

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For cellular viability assays, cells were seeded in 384-well opaque white tissue culture and assay plates (Corning) at 500 cells/well. 12–18 hours after seeding, cells were treated with compounds at the indicated concentrations for 48 hours. Cellular ATP levels were quantified using CellTiter-Glo Luminescence Assay (Promega) on an Envision multi-plate reader (PerkinElmer). Relative viability was normalized to the respective untreated condition of each cell line using RStudio and plotted in PRISM 9 (GraphPad software). For data presentation, the mean and standard deviation (s.d.) for the four or six biological replicates of each data point in a representative experiment is presented. Sigmoidal non-linear regression models were used to compute the regression fit curves.

Wang F., Graham E.T., Naowarojna N., Shi Z., Wang Y., Xie G., Zhou L., Salmon W., Jia J.M., Wang X., Huang Y., Schreiber S.L, & Zou Y. (2021). PALP: a rapid imaging technique for stratifying ferroptosis sensitivity in normal and tumor tissues in situ. Cell chemical biology, 29(1), 157-170.e6.

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Cell Viability Luminescence Assay

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Cell viability was measured using the CellTiter-Glo luminescence assay (Promega, UK), in accordance with the manufacturer׳s instructions.

Olayanju A., Copple I.M., Bryan H.K., Edge G.T., Sison R.L., Wong M.W., Lai Z.Q., Lin Z.X., Dunn K., Sanderson C.M., Alghanem A.F., Cross M.J., Ellis E.C., Ingelman-Sundberg M., Malik H.Z., Kitteringham N.R., Goldring C.E, & Park B.K. (2015). Brusatol provokes a rapid and transient inhibition of Nrf2 signaling and sensitizes mammalian cells to chemical toxicity—implications for therapeutic targeting of Nrf2. Free Radical Biology & Medicine, 78, 202-212.

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Cell Viability Assay for Compound Screening

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Cells were seeded onto 384-well tissue culture treated plates at a density of 25,000 cells/ml. 96-well drug plates were created such that each plate contained one compound serially diluted 1:2 at dose range specified with DMSO controls and compounds were transferred robotically. After treatment with a single compound (single agent IC₅₀) or combination of compounds (synergy studies) at serial two-fold dilutions, plated cells were analyzed for cell viability at the time points specified (zero to five days) using the Cell-TiterGlo luminescence assay (Promega) per the manufacturer’s instructions. Luminescence was read on a Fluostar Omega Reader (BMG Labtech). IC₅₀ values were calculated from ATP luminescence measurements using log-transformed, normalized data in GraphPad Prism 5.0 (GraphPad Software, Inc.).

Guenther L.M., Dharia N.V., Ross L., Conway A., Robichaud A.L., Catlett II J.L., Wechsler C.S., Frank E.S., Goodale A., Church A.J., Tseng Y.Y., Guha R., McKnight C.G., Janeway K.A., Boehm J.S., Mora J., Davis M.I., Alexe G., Piccioni F, & Stegmaier K. (2018). A combination CDK4/6 and IGF1R inhibitor strategy for Ewing sarcoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(4), 1343-1357.

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