ONE-Glo™ EX Luciferase Assay System

High- or ultrahigh-throughput quantitation of firefly luciferase expression in mammalian cells is commonly performed by measuring luminescence from 96-, 384- or 1,536-well plates. The ONE-Glo™ EX Luciferase Assay System provides both the high sensitivity and long-lived luminescence required to batch-process multiple plates in these assay formats.

Assay Advantages

• Increased stability of reconstituted reagent at room temperature or 4°C simplifies repeat use over long experiments and reduces waste; no odor-causing compounds in the reagent.
• Bright and stable signal that can be measured for hours enables more consistent luminescence readings; ideal for screening and batch processing.
• The homogenous firefly luciferase detection reagent can be used in both single- or dual-luciferase assays; compatible with any plate size up to 1,536-well format.
• Fewer Unwanted Effects from Sample Components. Reduced sensitivity to culture media, phenol red and luciferase inhibitors compared to other luciferase assays.

Stable Luminescence Signal Supports Batch Processing

The ONE-Glo™ EX Assay offers many of the same benefits of the ONE-Glo™ Assay, including use of  5´-fluoroluciferin substrate and an add-mix-read, or homogeneous, protocol. However, the ONE-Glo™ EX Reagent uses a new assay chemistry to increase the stability of both the luminescence signal and the reconstituted reagent. The approximately 2-hour signal half-life provides greater flexibility in assay design.

The example data shown here illustrate how the ONE-Glo™ EX signal persisted over 2 hours. In this experiment, 100µl purified firefly luciferase (13.8ng/ml) was combined in a 96-well plate with 100µl of Bright-Glo™, ONE-Glo™, ONE-Glo™ EX, Dual-Glo® Luciferase or Steady-Glo® Reagents. Luminescence was measured periodically over 2 hours, n = 8.

More Storage Options for Reconstituted Reagent

The reconstituted ONE-Glo™ EX reagent can be stored at room temperature for longer periods, so there is less variability in reagent performance during long experiments or screens and less sample waste. ONE-Glo™ EX Reagent is the firefly luciferase detection reagent used in the Nano-Glo® Dual-Luciferase® Reporter (NanoDLR™) Assay System; the same reagent is used for single- or dual-luciferase assays.

This data shows the stability of the ONE-Glo™ EX Reagent over 20 days when stored at 22°C or 7°C.