Dual-Luciferase® Reporter Assay System Technical Manual

Literature # TM040

The Dual-Luciferase® Reporter (DLR™) Assay System provides an efficient means of performing dual reporter assays. In the DLR™ Assay, the activities of firefly (Photinus pyralis) and Renilla (Renilla reniformis, also known as sea pansy)luciferases are measured sequentially from a single sample. The firefly luciferase reporter is measured first by adding Luciferase Assay Reagent II (LAR II) to generate a “glow-type” luminescent signal. After quantifying the firefly luminescence, this reaction is quenched, and the Renilla luciferase reaction is initiated by simultaneously adding Stop & Glo® Reagent to the same tube. The Stop & Glo® Reagent also produces a “glow-type” signal from the Renilla luciferase, which decays slowly over the course of the measurement. In the DLR™ Assay System, both reporters yield linear assays with subattomole (<10^-18)sensitivities and no endogenous activity of either reporter in the experimental host cells. Furthermore, the integrated format of the DLR™ Assay provides rapid quantitation of both reporters either in transfected cells or in cell-free transcription/translation reactions.
Promega has two series of firefly and Renilla luciferase vectors, pGL4 and pRL, designed for use with the DLR™ Assay Systems. These vectors may be used to co-transfect mammalian cells with any experimental and control reporter genes.