Celltiter glo

Manufactured by PerkinElmer

Sourced in United States

CellTiter-Glo is a cell viability assay that quantifies the amount of ATP present in metabolically active cells. It is a homogeneous, luminescent-based method that determines the number of viable cells in a culture. The assay principle is based on the luciferase reaction, which generates a stable luminescent signal proportional to the amount of ATP present in the sample.

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Lab products found in correlation

Celltiter glo, by Promega (15 mentions) Victor 1420 multilabel counter, by PerkinElmer (4 mentions) Viewlux, by PerkinElmer (3 mentions) Envision plate reader, by PerkinElmer (2 mentions) Sodium iodoacetate, by Merck Group (1 mentions) Envision 2105 multimode plate reader, by PerkinElmer (1 mentions) Oligomycin a, by Merck Group (1 mentions) Victor x3 multimode plate reader, by PerkinElmer (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Ensight multimodal plate reader, by PerkinElmer (1 mentions) Enspire multimode plate reader, by PerkinElmer (1 mentions) Envision 2104 multilabel reader, by PerkinElmer (1 mentions) Envision multilabel reader, by PerkinElmer (1 mentions) 1536 well black clear bottom plates, by Greiner (1 mentions) Celltiter glo reagent, by Promega (1 mentions) Megafasl, by Adipogen (1 mentions) Victor x5, by PerkinElmer (1 mentions) Luciferin reagent, by Promega (1 mentions) 1536 well plates, by Greiner (1 mentions) G7572, by Promega (1 mentions)

Spelling variants of this product

CellTiter-Glo assay (4 citations) CellTiter-Glo® Reagent (2 citations) CellTiter-Glo Luminescent Cell Viability Assay (2 citations)

21 protocols using celltiter glo

Evaluating Combination Therapy Efficacy

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Control (DMSO), Olaparib (No.S1060, Selleckchem) (25 μM.mL-1), OXA (10 uM.mL-1), cis Platinum (5 μM.mL-1) and CDK1 inhibitor (AG-024322, BIOQUOTR, and 837364-57-5) (0.12 μM.mL-1) were added to cells (500/well) in a 6-well plate. After 2 weeks of culturing, the formation of colonies or colosphere was evidently visible, the cell colonies were then fixed, stained with 0.1% crystal violet in 20% methanol solution, and counted. This process was repeated three times per solution type.
In a 96-well transparent bottom blackboard, 3,000 cells were planted in each well (organoids were planted in Matrigel). The drug was then added to each well according to a 10-fold concentration gradient. Cell viability as determined by Adenosine triphosphate (ATP) levels (Promega, Madison, WI, United States) were assayed by CellTiter-Glo using a luminometer (PerkinElmer Life and Analytical Sciences, Boston, MA, United States) 48 h later.

Li H., Wang C., Lan L., Wu W., Evans I., Ruiz E.J., Yan L., Zhou Z., Oliveira J.M., Reis R.L., Hu Z., Chen W., Behrens A., He Y, & Zhang C. (2021). PARP1 Inhibitor Combined With Oxaliplatin Efficiently Suppresses Oxaliplatin Resistance in Gastric Cancer-Derived Organoids via Homologous Recombination and the Base Excision Repair Pathway. Frontiers in Cell and Developmental Biology, 9, 719192.

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Inhibitor Pretreatment and Apoptosis Assay

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Cells were plated into sterile Nunc Edge 96-well tissue culture-treated plates at a density of 10,000 cells/well and preincubated (or chased) with 20 µM of the indicated inhibitor or DMSO vehicle for the indicated time before (or after) apoptotic induction. 3 h after induction, cellular viability was measured using CellTiter-Glo® on a PerkinElmer EnVision plate reader.

Solania A., González-Páez G.E, & Wolan D.W. (2019). A selective and rapid cell-permeable inhibitor of human caspase-3. ACS chemical biology, 14(11), 2463-2470.

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Quantifying Cellular Energy Metabolism

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The ATP cell content was measured using Cell Titer Glo (#G7571, Promega, Charbonnières Les Bains, France). In summary, the cells were seeded at 2000 cells/well in three replicates in 384-well ViewPlates (Perking Elmer). After an overnight, oligomycin A (Sigma-Aldrich) and sodium iodoacetate (Sigma-Aldrich) were added, either both or in combination. Following a 1 h incubation, the cell confluence was assessed with IncuCyte for further normalization and 40 μL of Cell Titer Glo reaction mix was added to each well, reaching a final volume of 80 μL. The plates were then analyzed for luminescence with an EnVision 2105 Multimode Plate Reader (Perkin Elmer). By comparing the different conditions, the global ATP and the percentages of both glycolytic and mitochondrial ATP were determined.

Luna-Yolba R., Marmoiton J., Gigo V., Marechal X., Boet E., Sahal A., Alet N., Abramovich I., Gottlieb E., Visentin V., Paillasse M.R, & Sarry J.E. (2021). Disrupting Mitochondrial Electron Transfer Chain Complex I Decreases Immune Checkpoints in Murine and Human Acute Myeloid Leukemic Cells. Cancers, 13(14), 3499.

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Cytotoxicity Evaluation of Oral Compounds

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The toxicity of the CPC, CHX, and mouthwashes was assessed with CellTiter-Glo (Promega, Madison, USA) following the manufacturer’s instructions. Briefly, MDCK or HeLa cells were seeded on white 96-well plates (1x10⁴ cells/plate) and treated with the aforementioned compounds at different concentrations. After 24 hours, 100 μL of CellTiter-Glo reagent was added to each well and the luminescence was measured using a Victor X3 Multimode Plate Reader (Perkin Elmer, Shelton, USA).

Rius-Salvador M., García-Múrria M.J., Rusu L., Bañó-Polo M., León R., Geller R., Mingarro I, & Martinez-Gil L. (2024). Cetylpyridinium chloride and chlorhexidine show antiviral activity against Influenza A virus and Respiratory Syncytial virus in vitro. PLOS ONE, 19(2), e0297291.

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Apoptosis and Cell Proliferation Assays

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Cells were stained using the Apotest FITC kit (Nexins Research, Kattendijke, The Netherlands). Briefly, cells were incubated with annexin V fluorescein isothiocyanate (0.2 μg/ml) on ice for 1 h. Propidium iodide (PI) (1.4 μg/ml) was added 5 min before cells were analyzed using an LSRII flow cytometer (BD Biosciences, San Jose, CA, USA). Cells negative for both annexin V and PI staining were considered viable. CellTiter-Glo (Promega, Madison, WI, USA) measures ATP levels in cells by use of luciferase and was used to determine relative cell proliferation. Cells were seeded in 96-well optical plates and treated as indicated. CellTiter-Glo reagent was added according to the manufacturer's protocol and luminescence was determined using Victor 1420 multilabel counter (PerkinElmer Inc., Waltham, MA, USA).

Olsen O.E., Wader K.F., Misund K., Våtsveen T.K., Rø T.B., Mylin A.K., Turesson I., Størdal B.F., Moen S.H., Standal T., Waage A., Sundan A, & Holien T. (2014). Bone morphogenetic protein-9 suppresses growth of myeloma cells by signaling through ALK2 but is inhibited by endoglin. Blood Cancer Journal, 4(3), e196-.

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Cell Viability Assay Protocol

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A CellTiterGlo (Promega) assay was performed to assess viability. A total of 30 µL of CellTiterGlo was added to 40 µL of standards or spheroids in media and incubated for 1 h at 37 °C and read on an EnSight Multimodal plate reader (Perkin Elmer).

Trossbach M., de Lucas Sanz M., Seashore-Ludlow B, & Joensson H.N. (2022). A Portable, Negative-Pressure Actuated, Dynamically Tunable Microfluidic Droplet Generator. Micromachines, 13(11), 1823.

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Cytotoxicity Assay of Drug Candidates

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MDA-MB-231, HCC1937 and HCC1806 were plated in black wall clear bottom 96-well plates at 10,000 cells/well and 5,000 cells/well 24 hrs before treatment, for 2 hr and 48 hr treatment, respectively. On the day of drug treatment, of SW43, SW IV-52s, Taxol or SW IV-134 were added at concentrations from 10 pM – 100 μM. Cells with appropriate treatments were incubated for either 2 or 48 hrs. For 2 hr treatments, drug solution was removed and cells were replenished with fresh media, and then allowed to regrow for 48 hrs. Viability was determined using commercially available CellTiter Glo® (Promega, Madison WI) chemiluminescent assay that measures ATP. For 48 hr treatment, the media was removed and CellTiter Glo® was added, then plates were immediately read on a Perkin Elmer Enspire® Multimode Plate Reader. Luminescence detected for each well was normalized to the untreated control and data was calculated as percent viability. All experiments were completed in triplicate at three independent times. Data was plotted using GraphPad Prism version 6.0 non-linear fit sigmoidal dose response variable slope and effective concentrations for 50% maximal reduction in cell viability (EC₅₀) were calculated.

Makvandi M., Tilahun E.D., Lieberman B.P., Anderson R.C., Zeng C., Xu K., Hou C., McDonald E.S., Pryma D.A, & Mach R.H. (2015). The sigma-2 receptor as a therapeutic target for drug delivery in triple negative breast cancer. Biochemical and biophysical research communications, 467(4), 1070-1075.

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Quantifying Glioma Cell Viability with ATA

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The CellTiterGlo® (Promega, San Luis Obispo, CA) assay was used to assess the cell viability after ATA treatment as previously described with minor modifications [50 (link)]. Briefly, T98G, A172, and GBM44 cells were seeded at a density of 3000 cells/well (80 μL) in 96 well plates. Increasing concentrations of ATA were added to the different wells (8 replicate wells for each condition) and incubated for 72 hr in 37°C, 5% CO₂ atmosphere. Subsequently, CellTiterGlo® reagent was added to each well and luminescence was measured using Perkin Elmer Envision 2104 Multilabel Reader. On all 96 well plates, wells containing vehicle only or the positive control compound MG132 (a proteasome inhibitor toxic to most cell lines at 2 μM) were also included. Raw values were normalized on a plate-by-plate basis such that 100% cell viability was equivalent to the mean of vehicle wells and 0% cell viability was equivalent to the mean of the MG132 positive control. The normalized data was used to assess viability of glioma cells after ATA treatment.

Roos A., Dhruv H.D., Mathews I.T., Inge L.J., Tuncali S., Hartman L.K., Chow D., Millard N., Yin H.H., Kloss J., Loftus J.C., Winkles J.A., Berens M.E, & Tran N.L. (2017). Identification of aurintricarboxylic acid as a selective inhibitor of the TWEAK-Fn14 signaling pathway in glioblastoma cells. Oncotarget, 8(7), 12234-12246.

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Cell Proliferation Assay with GLS Knockdown

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A bulk volume of cells was diluted to accommodate a seeding density of 2,000 cells/100μL (MDA-MB-231, MDA-MB-453, SUM-159PT) and 3,000 cells/100μL (SKBR3) in four 96-well white, clear bottom cell culture plates. To measure proliferation, each plate was incubated in the presence or absence of doxycycline and assayed with CellTiter-GLO (Promega) at 0, 3, 6, and 9 days after 3 days of doxycycline induction. On the day of analysis, a white plate backing was adhered to the bottom of each plate following microscopic examination. 100μL CellTiter-GLO reagent was added to each well and read in the Envision Multilabel Reader (Perkin Elmer). The unused cells were pelleted and frozen in -80°C for western analysis (1x10⁶ cells) and qRT-PCR (100,000 cells) to confirm knockdown. The metabolic rescue of GLS knockdown required adaptation of the cell proliferation assay procedure by adding 4mM Dimethyl 2-Oxoglutarate to the growth media of doxycycline induced GLS shRNA stable cell lines.

Lampa M., Arlt H., He T., Ospina B., Reeves J., Zhang B., Murtie J., Deng G., Barberis C., Hoffmann D., Cheng H., Pollard J., Winter C., Richon V., Garcia-Escheverria C., Adrian F., Wiederschain D, & Srinivasan L. (2017). Glutaminase is essential for the growth of triple-negative breast cancer cells with a deregulated glutamine metabolism pathway and its suppression synergizes with mTOR inhibition. PLoS ONE, 12(9), e0185092.

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High-Throughput Cell Viability Assay

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Cells in growth medium were plated into a 1536 well plate (5 μL/well; 250 cells/well) using a GNF Bottle Valve liquid handler. A Labcyte Echo acoustic transfer instrument was used to transfer 15 nL of compounds in DMSO to each well (final concentration 30 μM, 9.5 μM, 3 μM, 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, and 0.01 μM). The cells were then incubated (37 °C, 95% Humidity, 5% CO₂) for 3 days and 6 hours prior to addition of 4 μL of 50% Cell-Titer Glo (Promega) in water using a GNF Bottle Valve liquid handler. Plates were incubated with Cell Titer Glo for 15 minutes at room temperature prior to reading luminescence (5 s exposure) on a Perkin Elmer ViewLux. For determining GI50 values, data was normalized to a day 0 cell count measured using a cell plate copy that was not treated with compound and growth inhibition dose-response curves were calculated using Helios.

Isobe Y., Okumura M., McGregor L.M., Brittain S.M., Jones M.D., Liang X., White R., Forrester W., McKenna J.M., Tallarico J.A., Schirle M., Maimone T.J, & Nomura D.K. (2020). Manumycin Polyketides Act as Molecular Glues Between UBR7 and P53. Nature chemical biology, 16(11), 1189-1198.

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