LB 9507 Chapter 9: Dual-Luciferase
TM
Reporter Gene Assay
9-1
9. Dual-Luciferase
TM
Reporter Gene Assay
The Dual-Luciferase
TM
1
Reporter Assay System
2
from Promega
offers the exceptional speed, sensitivity and convenience of two
luciferase reporter assays in a single-tube format. The Dual-
Luciferase
TM
Reporter Gene Assay protocol of the Lumat LB 9507
is designed for automatic measurement of this assay. This assay
is performed in the following way: after injection of the first reagent
(substrate luciferin for firefly luciferase) the light emission is
measured. Then the second reagent is injected (Stop and Glo
TM 3
reagent). This quenches the luminescence signal from the firefly
reaction. The simultaneous activation of the second luciferase
from Renilla reniformis
4
is also achieved. In the second measuring
step the light emission of the Renilla luciferase
5
is detected.
1
Dual-Luciferase is a trademark of Promega Corp.
2
U.S. Pat. No. 5,744,320 has been issued to Promega Corporation for quenching reagents and
assays for enzyme mediated luminescence.
U.S. Pat. No. 5,283,179, 5,641,641 and 5,650,289 have been issued to Promega Corporation for a
firefly luciferase assay method which affords greater light output with improved kinetics as
compared to the conventional assay.
3
Stop and Glo is a trademark of Promega Corp.
4
The cDNA encoding luciferase from Renilla reniformis is covered by U.S. Pat. No. 5,292,658
assigned to the University of Georgia Research Foundation Inc., and sublicensed by Promega
Corporation from SeaLite Sciences, Inc., Norcross, GA. The pRL family of Renilla luciferase cDNA
vectors is for research use only.
5
as above
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