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Dual-Luciferase® Reporter 1000 Assay System Technical Manual

Instructions for Use of Product(s)

Literature # TM046

Dual-Luciferase® Reporter (DLR™) Assay System provides an efficient means of performing dual reporter assays. In the DLR™ Reporter 1000 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 stabilized 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 stabilized signal from the Renilla luciferase, which decays slowly over the course of the measurement. In the DLR™ 1000 Assay System, both reporters yield linear assays with subattomole sensitivities and no endogenous activity of either reporter in the experimental host cells. Furthermore, the integrated format of the DLR™ 1000 Assay provides rapid quantitation of both reporters either in transfected cells or in cell-free transcription/translation reactions.

The DLR™ 1000 Assay System was developed for larger volume users of the DLR™ Assay and, in particular, is configured for use in 96-well luminometry plates. Additional volume of both assay reagents is supplied to allow for priming of reagent injectors. Sufficient lysis reagent (Passive Lysis Buffer, PLB) has been supplied to allow for addition of 20µl per well in 96-well plates. For applications requiring more lysis reagent (e.g., >100µl/well), additional PLB may be purchased separately (Cat.# E1941). The components of the DLR™ 1000 Assay System are identical in formulation to those provided with the DLR™ Assay System (Cat.# E1910 and E1960).

Promega has several series of firefly and Renilla luciferase vectors, pGL4, pGL3 and pRL,designed for use with the DLR™ Assay Systems. The vectors may be used to co-transfect mammalian cells with any experimental and control reporter genes.

Summary of Changes
The following change was made to the 11/18 revision of this document:
1.The document design was updated.
2. Patent information was updated.
3. Additional GloMax® Instruments were added to the list of available products.

Revised 11/18.

Experienced User Protocols