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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.
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.
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Protocols
Complete Protocol
ONE-Glo™ EX Luciferase Assay System Technical Manual
PDF (756 KB)
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Certificate of Analysis
Lookup Certificate of AnalysisStorage Conditions
-30C TO -10C
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.