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Abstract for Dual-Luciferase® Reporter 1000 Assay System
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 “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™ 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 many series of firefly and Renilla luciferase vectors,
pGL4, pGL3, phRL 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.
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