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The Dual-Luciferase Reporter (DLR) Assay System provides an efficient means of performing two reporter assays. The activities of firefly (Photinus pyralis) and Renilla (Renilla reniformis or sea pansy) luciferases are measured sequentially from a single sample. The firefly luciferase reporter is measured first by adding Luciferase Assay Reagent II to generate a luminescent signal lasting at least one minute. After quantifying the firefly luminescence, this reaction is quenched, and the Renilla luciferase reaction is initiated simultaneously by adding Stop & Glo Reagent to the same sample. Both assays can be completed in about 4 seconds using a luminometer with reagent auto-injectors. Both reporters yield linear assays with attomole (<10(-18)) sensitivities and no endogenous activity in the experimental host cells. The integrated format of the DLR Assay provides rapid quantitation of both reporters either in transfected cells or in cell-free transcription/translation reactions.
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