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This product is available through the Promega Helix onsite stocking program in a –20°C Helix Freezer. The program offers numerous convenient solutions to meet your lab's needs. Helix Freezers are available in two sizes: 5.7 cubic ft. or 9.7 cubic ft.
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The ONE-Glo™ + Tox Assay combines luciferase assay chemistry with a cell viability marker to better understand reporter gene expression in the context of cell health. The assay uses a two-step, addition-only process to make these measurements in a single well of a plate, negating the need to run parallel assays. The simple "add-mix-read" assay format and optimized reagents combine to create a flexible, automation-friendly assay that can be scaled to meet your needs.
In the first part of the assay, a nonlytic fluorescence-based method (CellTiter-Fluor™ Cell Viability Assay) is used to measure the relative number of live cells in culture. The CellTiter-Fluor™ Assay measures a conserved and constitutive protease activity within living cells. This live-cell protease activity is restricted to intact viable cells and is measured using a fluorogenic, cell-permeant peptide substrate (glycylphenylalanyl-aminofluorocoumarin; GF-AFC). The substrate enters intact cells where it is cleaved by the live-cell protease to generate a fluorescent signal proportional to the number of living cells. The live-cell protease becomes inactive upon loss of cell membrane integrity and leakage into the surrounding culture medium. Fluorescence of the free AFC fluorophore is measured with a microplate reader or CCD imager using an excitation wavelength of 380–400nm and emission wavelength of 505nm.
The second part of the assay uses the ONE-Glo™ Luciferase Assay System to quantify firefly luciferase gene expression from cells expressing the reporter. Ideally suited for high- and ultrahigh-throughput applications, the ONE-Glo™ Assay Reagent contains a fluoroluciferin substrate that gives increased stability and greater tolerance of sample components, and has less odor than standard luciferase assay reagents. Luminescence is measured with a microplate reader or CCD imager.
Niles, A.L. et al. (2007) A homogeneous assay to measure live and dead cells in the same sample by detecting different protease markers. Anal. Biochem. 366, 197–206.
In the experiment shown here, 1 × 104 (96-well plate; Panel A) or 5 × 103 (384-well plate; Panel B) cells expressing NFAT response element were treated with serial titrations of ionomycin in the presence of PMA for 6 hours. At specific concentrations, ionomycin and PMA work cooperatively to stimulate NFAT-dependent gene expression. However, higher concentrations of ionomycin result in cytotoxicity seen as a decrease in viability (fluorescence). A decrease in reporter expression (luciferase) activity is also observed due to the increase in cytotoxicity.
ONE-Glo™ + Tox Luciferase Reporter and Cell Viability Assay Technical Manual
PDF (524 KB)
ONE-Glo™ + Tox Luciferase Reporter and Cell Viability Assay FB129
PDF (124 KB)
ONE-Glo™ Luciferase Assay Buffer
ONE-Glo™ Luciferase Assay Substrate
PDF (0 B)
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U.S. Pat. Nos. 7,416,854, 7,553,632 and other patents pending.
Certain applications of this product may require licenses from others.
Improved ONE-Glo™ Assay. Higher sensitivity, long-lived luminescence, more storage options.
E8110, E8120, E8130, E8150
A bioluminescent method to kinetically monitor viability in cell culture up to 72 hours.
G9711, G9712, G9713
Measure viability, cytotoxicity and apoptosis in one sample well to confirm mechanism of cell death.
Ultra-sensitive detection of firefly and NanoLuc® luciferase activities in a single sample.
N1610, N1620, N1630, N1650, N1521, N1531, N1541, N1551
High-efficiency, low-toxicity transfection of many cell types, including iPS cells.
High-performance multimode plate reader for detecting luminescence, fluorescence and absorbance.
Enhanced performance in a range of cell types, including difficult-to-transfect cell lines.
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