The RealTime-Glo™ MT Cell Viability Assay is a non lytic, homogeneous, bioluminescent method to measure cell viability in real time using a simple, plate-based method. RealTime-Glo™ MT Cell Viability Assay determines the number of viable cells in culture by measuring the reducing potential of cells and thus metabolism (MT).

• Save time, cell culture and reagent costs
• Rapidly detect changes in cell viability
• Monitor cell viability in real time

The non lytic nature and rapid response of the assay make it possible to monitor cell viability over time in the same well, and enables easy multiplexing with other cell-based assays or downstream applications. The assay measures the reducing potential of viable cells and is ATP-independent, providing an orthogonal method for viability or cytotoxicity determination.

The assay involves adding NanoLuc® luciferase and a cell-permeant prosubstrate, the MT Cell Viability Substrate, to cells in culture. Viable cells reduce the proprietary prosubstrate to generate a substrate for NanoLuc® luciferase. This substrate diffuses from cells into the surrounding culture medium, where it is rapidly used by the NanoLuc® Enzyme to produce a luminescent signal. The signal correlates with the number of viable cells, making the assay well suited for cytotoxicity studies. Both the MT Cell Viability Substrate and NanoLuc® enzyme are stable in complete cell culture medium at 37 °C for at least 72 hours. No cell washing, removal of medium or further reagent addition is required to determine the number of viable cells.

Viable cells reduce the proprietary pro-substrate to generate a substrate for NanoLuc luciferase. This substrate diffuses from cells into the surrounding culture medium where it is rapidly used by the NanoLuc enzyme to produce a luminescent signal. The signal correlates with the number of viable cells, making the assay well suited for cytotoxicity studies. The reagent is stable and nontoxic to cells for up to 72 hours. No cell washing, removal of medium, or further reagent addition is required to determine the number of viable cells. The non-lytic nature of this assay enables cells to be monitored over time in the same well, which reduces the amount of cells used and cell culture costs, and in downstream applications, including assay multiplexing and nucleic acid analysis.