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Cancer Lett. 356, 994-1006. Lobatin B inhibits NPM/ALK and NF-κB attenuating anaplastic-large-cell-lymphomagenesis and lymphendothelial tumour intravasation. 2015

Kiss, I., et al.

Notes: SR-786 NPM/ALK-positive human anaplastic large cell lymphoma cell line was treated with lobatin B, a plant-derived natural compound. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and reversed transcribed into cDNA with the GoScript™ Reverse Transcription System. Levels of NPM/ALK and NF-κB expression was measured with the dye-based GoTaq® qPCR System. SR-786 cells were also analyzed for functioning metabolism with the CellTiter-Blue® Cell Viability Assay and apoptosis with the Apo-ONE™ Homogeneous Caspase-3/7 Assay. (4604)

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PLos ONE 10, e0129058. New small molecules targeting apoptosis and cell viability in osteosarcoma.  2015

Maugg, D. et al.

Notes: A phenotypic screen of a 25,000 compound library for inhibitors of U2OS osteosarcoma cell line growth identified two compounds that showed activity of osteosarcomas and not HepG2 or primary human osteoblasts.  Cell Viability in the primary and secondary screens was monitored with the CellTiter-Blue® Cell Viability Assay.  Subsequent studies identified the mechanism of cell death as apoptosis as judged by the Caspase-Glo® 3/7 Assay and cytotoxicity measured with the CellTox™ Green Cytotoxicity Assay.

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Int. J. Environ. Res. Public Health 12, 3915–3925. Evaluation of e-cigarette liquid vapor and mainstream cigarette smoke after direct exposure of primary human bronchial epithelial cells. 2015

Scheffler, S., Dieken, H., Krischenowski, O., Förster, C., Branscheid, D. and Aufderheide, M.

Notes: Primary normal human bronchial epithelial (NHBE) cells were cultivated on semi-permeable membranes of cell culture inserts exposed directly to vapor from 200 puffs of two different e-cigarette liquids (0% and 2.4% nicotine). Twenty-four hours post exposure, two assays were multiplexed to assess oxidative stress (ROS-Glo™ H2O2 Assay) and cell viability (CellTiter-Blue® Cell Viability Assay). First, the cells were incubated with 200µl of medium + 50µl of H2O2 Substrate Solution for 3 hours and 75µl of the mixture transferred to a white 96-well plate. An equal volume of Detection Solution was added, incubated for 20 minutes at room temperature and luminescence detected. The exposed cells had the remaining medium + H2O2 Substrate Solution removed and replaced with 300µl of fresh medium + 60µl of CellTiter-Blue® Reagent. After a 2-hour incubation, 100µl of solution was transferred to a black 96-well plate and fluorescence measured at 544nmEx/590nmEm. (4569)

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Current Chemical Genomics 3, 33-41. In vitro viability and cytotoxicity testing and same-well multi-parametric combinations for high-throughput screening 2009

Niles, A.L., Moravec, R.A. and Riss, T.L.

Notes: The authors review the use of in vitro cytotoxicity testing in drug discovery to characterize the toxic potential of new chemical entities (nce) at the earliest stages of profiling. DOI: 10.2174/1875397300903010033 (4000)

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Toxicology in Vitro 23, 1170-1171. Multiplexed assay panel of cytotoxicity in HK-2 cells for detection of renal proximal tubule injury potential of compounds 2009

Wu, Y., Connors, D., Barber, L., Jayachandra, S., Hanumegowda, U.M. and Adams, S.P.

Notes: The authors describe a multiplexed in vitro assay to detect nephrotoxicity and gain information about mechanism of cell death in HK-2 (human kidney-2) cells. The multiplexed assay involved an LDH assay to detect necrosis, a caspase-3/7 assay to detect apoptosis, a reazurin assay to assess metabolic state, and a DNA dye staining assay to monitor nuclear morphology. (4002)

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Mol. Cell. Endocrinol. 264, 50-60. Novel estrogen receptor beta transcript variants identified in human breast cancer cells affect cell growth and apoptosis of COS-1 cells. 2007

Treeck, O., Pfeiler ,G., Horn, F., Federhofer, B., Houlihan, H., Vollmer, A., and Ortmann, O.

Notes: This study identified two novel transcript variants of the estrogen receptor ERβ that were expressed in the ERα-negative breast cancer cell line MDA-MD-231. These variants were identified after amplification of ERβ transcripts from the breast cancer cell line by RT-PCR. The amplification products were then excised from gels and subcloned into the pTARGET™ Mammalian Expression Vector prior to sequencing. COS1 cells, which do not express the estrogen receptor, were then stably transfected with full-length ERβ or one of the splice variants and the effects on cell proliferation, apoptosis, and estrogen response were evaluated. In COS1 cells expressing either ERβ or the transcript variants cell proliferation decreased and basal apoptosis (caspase 3/7 activity) increased, compared to cells transfected with vector alone. Exposure to therapeutic doses of tamoxifen induced apoptosis in cells expressing the full-length ERβ but not in cells expressing either of the variant isoforms. (3618)

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FEBS J. 273, 2206–2222. Differing molecular mechanisms appear to underlie early toxicity of prefibrillar HypF-N aggregates to different cell types. 2006

Cecchi, C., Pensalfini, A., Baglioni, S., Fiorillo, C., Caporale, R., Formigli, L., Liguri, G., and Stefani, M.

Notes: The CellTiter-Blue® Cell Viability Assay was used to monitor viability of Hend murine endothelial cells and IMR90 fibroblasts in the presence of various concentrations (0.02, 0.2, 2.0 or 20 µm) of the N-terminal domain of the prokaryotic hydrogenase maturation factor (HypF-N). The cells were exposed to HypF-N for up to 24 hours before the cells were washed and cultured in fresh media. The CellTiter-Blue® Reagent was added and the cultures incubated for 1 hour before fluorescence readings were taken. Data was expressed as percent viability compared to a control. (3476)

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J. Biol. Chem. 281, 324-333. Mitochondrial transcription factor A induction by redox activation of nuclear respiratory factor 1. 2006

Piantadosi C.A., and Suliman, H.B.

Notes: Cell proliferation and viability of H4IIE rat hepatoma cells or HCC1937 human mammary primal ductal gland carcinoma cells were assessed with the CellTiter-Blue® Cell Viability Assay. Both cell lines were also exposed to 15µM of the lipophilic oxidant, tertiary butyl hydroperoxide (t-BOOH) for 5 hours. Incubations with the CellTiter-Blue® Cell Viability dye were carried out for 4 hours. Data are expressed as percent viability compared to control cultures. (3478)

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Nature 434, 652-658. Cyclophilin D-dependent mitochondrial permeability transition regulates some necrotic but not apoptotic cell death. 2005

Nakagawa, T., Shimizu, S., Watanabe, T., Yamaguchi, O., Otsu, K., Yamagata, H., Inohara, H., Kubo, T., Tsujimoto, Y.

Notes: In this study, the role of Cyclophilin-D (CypD) in the mitochondrial permeability transition (mPT) response was investigated using CypD-deficient mice. The CellTiter™ Blue Cell Viability Assay was used to measure the viability of mouse embryonic fibroblasts (MEF) and hepatocytes isolated from normal and CypD-deficient mice after exposure to various apoptotic stimuli and H2O2. (3398)

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Hum. Mol. Genet. 13(20), 2409-20. Correction of aberrant FGFR1 alternative RNA splicing through targeting of intronic regulatory elements 2004

Bruno, I.G., Jin, W., Cote, G.J.

Notes: Human U251 glioblastoma cell lines treated with antisense morpholino oligonucleotides were assessed for viability and apoptosis by multiplexing the CellTiter-Blue® Cell Viability and Apo-ONE® Homogeneous Caspase-3/7 Assays on single cell cultures. Cell viability was measured 4 hours after the addition of the CellTiter-Blue® Cell Viability Reagent to the cultures. Next, apoptosis measurements were performed on the same cell cultures by adding Apo-ONE® Homogeneous Caspase-3/7 Assay reagent to the cultures. Caspase-3/7 activity was then measured 12 hours later. (3168)

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Assay Drug Dev. Technol. 3, 469-477. HTS Compatible Assay for Antioxidative Agents Using Primary Cultured Hepatocytes 2003

Gaunitz, F. and Heise, K.

Notes: Primary hepatocytes from Sprauge-Dawley rats were incubated with various concentrations of tert-butylhydroperoxide to induce oxidative stress and then cell viability, cytotoxicity and apoptosis were assayed with Promega's CellTiter 96 AQueous One Solution Cell Proliferation Assay, CellTiter-Blue Cell Viability Assay, CellTiter-Glo Luminescent Cell Viability Assay, CytoTox-ONE Homogeneous Membrane Integrity Assay, Apo-ONE Homogeneous Caspase-3/7 Assay and assay systems. (2969)

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