The Dual-Luciferase? Reporter (DLR?) Assay System provides an efficient means of performing two reporter assays. In the DLR? Assay, 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 (LAR 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. In the DLR? Assay System, both reporters yield linear assays with attomole (<10–18) sensitivities and no endogenous activity 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.
For best results with the Dual-Luciferase? Assay, we recommend using a luminometer that has been validated for use with the assay. These luminometers are qualified as DLReady?. For a listing of qualified instruments, please visit the DLReady? Validated Luminometers page.
The pGL4 Luciferase Reporter Vectors are designed for use with the DLR? Assay Systems. A?Renilla?luciferase vector with constitutive expression may be used in combination with any experimental firefly luciferase vector to co-transfect mammalian cells.
Notice for Cat.# E1960 and E1980:?Sufficient Passive Lysis Buffer is provided to perform 1,000 assays with cells grown in 96-well plates (typically 20μl of 1X PLB per well). For applications requiring more lysis reagent (e.g., >100μl/well), additional Passive Lysis Buffer may be purchased separately.