Promega has launched the One-Glo + Tox assay, which is designed to improve the analysis of reporter gene expression in the context of cell health.
The assay combines luciferase chemistry with a cell viability marker in a two-step, addition-only process, to enable the measurement of cell viability and gene expression in a single well of a plate, negating the need to run parallel assays.
By analysing reporter activity in relation to cell viability, false results owing to toxicity or changes in cell number can be uncovered.
The first part of the assay is a non-lytic fluorescence assay (Celltiter-Fluor cell viability assay) that measures the relative number of the live cells in a culture population after experimental manipulation.
The Celltiter-Fluor assay measures a conserved and constitutive protease activity within live cells and therefore serves as a marker of cell viability.
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 second part of the assay uses the One-Glo Lucifease assay system to quantify firefly luciferase reporter gene expression from cells made to express this reporter enzyme.
Suitable for high- and ultra-high-throughput applications, the volumes of each assay component can be scaled to meet throughput needs and is amenable to automation up to a 1,536-well format.
The One-Glo + Tox assay contains a new fluoroluciferin substrate, resulting in a reagent that is more stable and tolerant to sample components.
Luminescence can be measured with a microplate reader or a CCD imager.
