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< GENERAL METHODS
Luminescence
Dual-luciferase quantitation in the
LMax
TM
microplate luminometer
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Molecular Devices MAXline Application Note #39
(2000).
Evelyn McGown, Ph.D. and Michael Su, M.S.
Summary. Colorimetric reporter assays
have been widely used, but biolumines-
cent reporter systems, especially using
firefly luciferase, are becoming increas-
ingly popular because of their speed,
sensitivity and wide dynamic range.
Promega Corporation has recently
introduced their Dual-Luciferase
®
Reporter Assay System which utilizes
the firefly (Photinus pyralis) and Renilla
(Renilla reniformis) luciferases. Because
the enzymes have different substrate
requirements, they can both be mea-
sured in a single reaction mixture. The
firefly luciferase is typically used as the
experimental reporter, and the renilla
luciferase serves as the control. Firefly
luciferase enzyme catalyzes the oxida-
tion of luciferin with the concomitant
release of light. The reaction requires
ATP, Mg
+2
and O
2
. Renilla luciferase
catalyzes the O
2
-dependent oxidation of
coelenterate luciferin (coelenterazine)
but does not require Mg or ATP. Both
reactions can easily be measured in
Molecular Devices' LMax microplate
luminometer. An example of results is
presented in this application note.
gene expression
Summary :
Promega Corporation has recently introduced their Dual-Luciferase ® Reporter Assay System which utilizes the firefly (Photinus pyralis) and Renilla (Renilla reniformis) luciferases. The firefly luciferase is typically used as the experimental reporter, and the renilla luciferase serves as the control.
Tags :
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