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Abstract for Dual-Luciferase® Reporter Assay System Technical Manual
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 three series of firefly and Renilla luciferase
vectors, pGL4, phRL 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.
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