Promega, E7120, ONE-Glo™ + Tox Luciferase Reporter and Cell Viability Assay

Name: ONE-Glo™ + Tox Luciferase Reporter and Cell Viability Assay
Brand: Promega
SKU: E7120

Sensitive Detection of Reporter Activity and Cell Health, Measures cell viability and firefly luciferase activity in the same assay well, Easy to use—simply add reagent, mix and read, Scalable to meet throughput needs, up to 1, 536-well format, Configuration: 10 plates

Catalog Number: E7120

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Category

Reagent Bottles

Pack Size1 vial

TagsLuciferase Assays Reporter Assays

Description

Understand Reporter Gene Activity in the Context of Cell Viability
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.
Step 1: Cell Viability Assay
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.
Step 2: ONE-Glo™ Luciferase Assay
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.
Reference
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.
Schematic of the ONE-Glo™+Tox Luciferase Reporter and Cell Viability Assay.

Specifications

{"type": "table", "rows": [{"name": "ONE-Glo™ Luciferase Assay Buffer", "conditions": {"Part #": "E605B", "Size": "1 × 100ml"}}, {"name": "ONE-Glo™ Luciferase Assay Substrate", "conditions": {"Part #": "E606B", "Size": "1 × 1 vial"}}, {"name": "GF-AFC Substrate", "conditions": {"Part #": "G608B", "Size": "2 × 50μl"}}, {"name": "Assay Buffer", "conditions": {"Part #": "G610B", "Size": "1 × 50ml"}}]}

Ordering

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