DLR assay system

The Dual-Luciferase® Reporter kit from Promega is a popular luminescent reporter assay to study gene transcription and regulation.

Dr Martin Mangold Dr Martin Mangold (7)

The Dual-Luciferase® Reporter (DLR™) assay system from Promega Corp. provides a luminescent reporter assay to study gene transcription and regulation. The activities of two luciferases are measured sequentially from a single sample reporting the transcription of the gene of interest and a transfection control.

Fig. 1: Explanation of DLR assay system

Both assays can be completed in about 4 seconds using a luminometer with reagent auto-injectors, such as in the Omega seriesVANTAstar®CLARIOstar®, and PHERAstar® FSX microplate readers. In the DLR™ assay system, both reporters yield linear assays with attomole 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.

Fig. 2: Dual Luciferase Reaction - Luciferase Assay Reagent II (LAR II) is injected in the first step and the Firefly reaction is started. Stop and Glo® buffer is injected in the second step, which quenches the Firefly reaction and initiates the Renilla reaction.

Using additional luciferases, with different emission peaks allow to set up huge luminescence multiplexing approaches using six different readouts. With the help of the CLARIOstar or VANTAstar and their luminescence spectral scanning feature, ideal combinations of different luciferase to be used in the same run can be identified.

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