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Celltiter glo viability assay

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The CellTiter-Glo viability assay is a luminescent cell-based assay that quantifies the amount of ATP present, which indicates the presence of metabolically active cells. The assay reagent causes cell lysis and generates a luminescent signal proportional to the amount of ATP present.

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Lab products found in correlation

Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Faststart sybr green, by Roche (2 mentions) First strand synthesis kit, by Thermo Fisher Scientific (2 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions) Rneasy kit, by Qiagen (2 mentions) Caspase glo 3 7 assay, by Promega (2 mentions) Dtx 880 multimode detector, by Beckman Coulter (2 mentions) Annexin 5 apoptosis detection kit apc, by Thermo Fisher Scientific (1 mentions) Victor x4, by PerkinElmer (1 mentions) Wst 1 assay, by Merck Group (1 mentions) Brdu elisa kit, by Merck Group (1 mentions) Abt 199, by Selleck Chemicals (1 mentions) A 1331852, by Selleck Chemicals (1 mentions) Accutase, by Promega (1 mentions) Guava easycyte, by Merck Group (1 mentions) Nheks, by Lonza (1 mentions) Singlequots supplements and growth factors, by Lonza (1 mentions) Nhdfs, by Lonza (1 mentions) Fgm 2 singlequots supplements, by Lonza (1 mentions) Kbm gold keratinocyte growth basal medium, by Lonza (1 mentions)

51 protocols using celltiter glo viability assay

1

CellTiter-Glo Viability Assay Protocol

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CellTiter-Glo viability assay (Promega) was performed by collecting cells at indicated time points and washing once with 1× PBS. A total of 50 μL cells in PBS were used per assay according to manufacturer instruction.
Burton E.M., Voyer J, & Gewurz B.E. (2022). Epstein–Barr virus latency programs dynamically sensitize B cells to ferroptosis. Proceedings of the National Academy of Sciences of the United States of America, 119(11), e2118300119.
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2

ATP-based Cell Viability and Apoptosis Assay

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ATP-based cell viability was determined using the CellTiter-Glo viability assay (Promega Corporation, Madison, WI, USA #G7573) and luminescence was measured using a Victor X4 (Perkin Elmer, Waltham, MA, USA). Apoptotic rate was quantified by staining cells with Annexin V and propidium iodide using a flow-cytometry commercial kit (eBioscience™ Annexin V Apoptosis Detection Kit APC, Waltham MA, USA, # 88-8007-74). Cells were analyzed by flow cytometry with a FACScan flow cytometer (Beckman Culture-Cytomics FC 500, Life Sciences Division, Indianapolis, USA) and FlowJo V10 (Tree Star LLC, Ashland, OR, USA) analytical software. Cellular DNA content was assessed by staining with propidium iodide (50 g/ml) and analyzed by flow cytometry. At least 20,000 events were acquired and all determinations were replicated at least twice.
Marchesini M., Gherli A., Montanaro A., Patrizi L., Sorrentino C., Pagliaro L., Rompietti C., Kitara S., Heit S., Olesen C.E., Møller J.V., Savi M., Bocchi L., Vilella R., Rizzi F., Baglione M., Rastelli G., Loiacono C., Starza R.L., Mecucci C., Stegmaier K., Aversa F., Stilli D., Winther A.M., Sportoletti P., Bublitz M., Dalby-Brown W, & Roti G. (2020). Blockade of Oncogenic NOTCH1 with the SERCA Inhibitor CAD204520 in T-cell Acute Lymphoblastic Leukemia. Cell chemical biology, 27(6), 678-697.e13.
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3

Colorectal Cancer Cell Cytotoxicity Assay

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TICs or the three human colorectal cancer cell lines HT29, SW480, and HCT116 (5 x 103 each) were cultured in 96-well flat bottom plates. Cells were exposed to increasing concentrations of salinomycin, 5-FU, or oxaliplatin (0.1 μM, 0.5 μM, 1 μM, 2 μM, 5 μM, and 10 μM) for 24–48 hours. The effect of the evaluated compounds on the cell number was assessed using the WST-1 assay (Sigma Aldrich) according to the manufaturer’s instructions. Cell proliferation was assessed using the bromodeoxyuridine (BrdU) ELISA kit (Sigma Aldrich), according to the manufacturer’s instructions. Additionally, cell numbers were analyzed applying the CellTiter-Glo Viability Assay (Promega). This assay is based on measuring a luminescence signal proportional to the ATP content present in healthy cells. The luminescence signal was quantified by a microplate luminometer as recommended by the manufacturer’s instructions.
Klose J., Trefz S., Wagner T., Steffen L., Preißendörfer Charrier A., Radhakrishnan P., Volz C., Schmidt T., Ulrich A., Dieter S.M., Ball C., Glimm H, & Schneider M. (2019). Salinomycin: Anti-tumor activity in a pre-clinical colorectal cancer model. PLoS ONE, 14(2), e0211916.
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4

Evaluating AML Cell Sensitivity to BCL-2 Inhibitors

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The sensitivity of AML cell lines towards ABT-199 (Selleck), A1331852 (Selleck) and S63845 (Appexbio) was assessed with CellTiter-Glo viability assay (Promega). Cells were seeded in a density of 1 × 105 cells/ml in a white 96-well plate. After 72 h of incubation 5 μl/well of CellTiter-Glo reagent was added and luminescence was measured with a Tecan Infinite M200 plate reader. Values were normalized to the untreated control sample.
Ewald L., Dittmann J., Vogler M, & Fulda S. (2019). Side-by-side comparison of BH3-mimetics identifies MCL-1 as a key therapeutic target in AML. Cell Death & Disease, 10(12), 917.
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5

Inducible YAP1/WWTR1 Expression

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YAP1- or WWTR1-expressing MKL-1 cells were treated with 2 μg/mL DOX for 3 and 6 days with complete refreshment on day 3. At each time point, cells were treated with Accutase to disrupt clumps, and suspensions were either plated in 96-well plates for a CellTiter-Glo viability assay (Promega) or lysed for SDS-PAGE.
Frost T.C., Gartin A.K., Liu M., Cheng J., Dharaneeswaran H., Keskin D.B., Wu C.J., Giobbie-Hurder A., Thakuria M, & DeCaprio J.A. (2023). YAP1 and WWTR1 expression inversely correlates with neuroendocrine markers in Merkel cell carcinoma. The Journal of Clinical Investigation, 133(5), e157171.
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6

Impact of Trametinib and Palbociclib on Tumor Secretome

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KPCmut tumor cells were treated with vehicle or combined trametinib (25nM) and palbociclib (500nM) for 6 days, and conditioned serum-free DMEM was collected as described above. Filtered conditioned media (CM) or basal serum-free media was then applied to various primary and established cell lines. To measure endothelial cell and fibroblast growth, 5,000 3B11 murine endothelial cells and 10,000 primary murine pancreatic fibroblasts were plated in 24-well dishes. Cells were counted daily in quadruplicate using a Guava Easycyte (Millipore) and media changed every 3 days. To analyze the impact on macrophage growth, 10,000 bone marrow-derived macrophages (BMDMs) were plated in 96 well dishes and, one day thereafter, were exposed to CM or basal serum-free media diluted 1:2 in DMEM containing 10% FBS and 10 ng/mL Csf1. Cell viability was assessed using the CellTiter-Glo Viability Assay (Promega) according to the manufacturer’s protocol.
Ruscetti M., Morris JP I.V., Mezzadra R., Russell J., Leibold J., Romesser P.B., Simon J., Kulick A., Ho Y.J., Fennell M., Li J., Norgard R.J., Wilkinson J.E., Alonso-Curbelo D., Sridharan R., Heller D.A., de Stanchina E., Stanger B.Z., Sherr C.J, & Lowe S.W. (2020). Senescence-Induced Vascular Remodeling Creates Therapeutic Vulnerabilities in Pancreas Cancer. Cell, 181(2), 424-441.e21.
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7

Cytocompatibility Evaluation of DA-ESCMs

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Normal adult human dermal fibroblasts (NHDFs, Lonza) were cultured in FBM-2 Basal Medium plus FGM-2 SingleQuots supplements (Lonza), normal adult human epidermal keratinocytes (NHEKs, Lonza) in KBM-Gold Keratinocyte Growth Basal Medium plus SingleQuots Supplements and Growth Factors (Lonza) and RAW 264.7 monocytes (ATCC, Manassas, VA, USA) in DMEM plus 10% FBS. All cells were seeded at 1 × 104 cells/cm2 in 24-well plates and cultured for 24 h at 37 °C and 5% CO2 before adding the experimental treatment. All media were supplemented with 500 IU/mL penicillin, 500 μg/mL streptomycin, and 2.5 μg/mL amphotericin-B. After overnight incubation, DA-ESCMs were placed in the wells. After 24 and 72 h, wells were imaged microscopically, and cell viability was quantified using the CellTiter-Glo® viability assay (Promega). Results were normalized as a percent viability versus cells grown with unloaded DA-ESCMs. Treatments were accepted as cytocompatible if they met or surpassed the 70% cytocompatibility minimum, as established by ISO 10993-5 [22 ].
Harrison Z., Montgomery E.C., Bush J.R., Gupta N., Bumgardner J.D., Fujiwara T., Baker D.L, & Jennings J.A. (2023). Cis-2-Decenoic Acid and Bupivacaine Delivered from Electrospun Chitosan Membranes Increase Cytokine Production in Dermal and Inflammatory Cell Lines. Pharmaceutics, 15(10), 2476.
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8

Antiviral Efficacy Screening of Compounds

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MDCK cells were seeded and grown for 18–24 h to a confluent monolayer in 96-well plate. The medium was changed to DF-12 medium containing 2 μg/mL TPCK-trypsin before PBS washing twice. Cells were infected with influenza virus at an MOI of 0.005 in DF-12 medium containing 2 μg/mL TPCK-trypsin in the presence of various concentrations (ranging from 0.005 μM to 100 μM by a three-fold dilution) of the test compound. After 72 h incubation at 37 °C in CO2 incubator, the antiviral activity of test compounds was measured using Celltiter-Glo viability assay (Promega). The concentration for 50% maximal effect (EC50) was calculated by Origin 8 software.
Zong K., Xu L., Hou Y., Zhang Q., Che J., Zhao L, & Li X. (2021). Virtual Screening and Molecular Dynamics Simulation Study of Influenza Polymerase PB2 Inhibitors. Molecules, 26(22), 6944.
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9

Characterization of Lung Cancer Cell Lines

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Mouse lung cancer-derived cell lines were generated as previously described (22 (link)) with recombination of the KrasLSL and Pik3caLat alleles verified by PCR (7 (link),18 (link)). H460 and A549 cells were obtained from the labs of Trever Bivona and Frank McCormick (UCSF), respectively, where authentication was recently performed. Cells were engineered to co-express luciferase and EGFP by infection with the lentiviral vector pLV430G-oFL-T2A-eGFP (23 (link)) and isolated using a FacsAria III (BD).
Proliferation was assessed by plating 1000 cells/well in 96-well plates and treating with the following agents: 1. DMSO control; 2. MEK1/2 inhibitor (PD0325901); 3. Pan-class 1 PI3′-kinase inhibitor (GDC-0941); 4. Selective PI3Kα inhibitor (BYL719); 5. AKT1-3 inhibitor (MK-2206) or various combinations as indicated. 72 hours after drug addition, a Cell-Titer-Glo viability assay was performed (Promega). Transition though S phase was assessed by incubating cells with 10μM BrdU for the final 3 hours of a 24-hour drug treatment. Cells were stained with anti-BrdU-FITC and quantified using a FACS-Calibur (BD).
Green S., Trejo C.L, & McMahon M. (2015). PIK3CAH1047R accelerates and enhances KRASG12D-driven lung tumorigenesis. Cancer research, 75(24), 5378-5391.
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10

Estrogen Receptor+ Breast Cancer Organoid Culture

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Organoids were derived using the human Tumor Dissociation Kit (Miltenyi Biotec) from the ER+ patient derived xenografts from the Breast Cancer Genome Guided Therapy Study according to a previously described protocol (15 (link)). Organoids were cultured in 96-well low binding NanoCulture plate (Organogenix) in DMEM supplemented with 10% FCS, 1% glutamax, 1% sodium pyruvate, non-essential amino acids, and 1% Penicillin-Streptomycin (Life Technologies) at 37°C, 5% CO2. Organoid growth was measured as previously described (15 (link)) using the CellTiter-Glo Viability assay (Promega).
Cairns J., Ingle J.N., Kalari K.R., Goetz M.P., Weinshilboum R.M., Gao H., Li H., Bari M.G, & Wang L. (2021). Anastrozole regulates fatty acid synthase in breast cancer. Molecular cancer therapeutics, 21(1), 206-216.
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