Description
Bright, Stable, Firefly Luciferase Assay with Improved Handling and Performance
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.
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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.
