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Abstract for Luciferase Assay System
The Luciferase Assay System is substantially improved over conventional
assay methods in both sensitivity and simplicity. Light is produced by
converting the chemical energy of luciferin oxidation through an electron
transition, forming the product molecule oxyluciferin. Firefly luciferase, a
monomeric 61kDa protein, catalyzes luciferin oxidation using ATP-Mg2+
as a cosubstrate. In the conventional assay for luciferase, a flash of light is
generated that decays rapidly after the enzyme and substrates are combined.
The Luciferase Assay System incorporates coenzyme A(CoA) for improved
kinetics, allowing greater enzymatic turnover resulting in increased light
intensity that is nearly constant for at least 1 minute. The Luciferase Assay
System yields linear results over at least eight orders of magnitude. Less than
10-20 moles of luciferase have been detected under optimal
conditions. Generally, 100-fold greater sensitivity can be achieved over the
chloramphenicol acetyltransferase (CAT) assay.
The Luciferase Assay System was developed for reporter quantitation in
mammalian cells. The Luciferase Assay System (Cat.# E1500), provided with Cell
Culture Lysis Reagent (CCLR), can also be used for reporter quantitation in
plant and bacterial cells; however, the Luciferase Assay System with Reporter
Lysis Buffer (Cat.# E4030) is not suitable for these applications.
The Luciferase Reporter 1000 Assay System (Cat.# E4550) was designed to meet
the needs of users who perform a large number of assays, particularly in 96-well
plates. The system contains sufficient reagents to perform 1,000 luciferase
assays (100µl per assay). For users working with transformed cells, a cell
lysis buffer will be needed for sample preparation prior to luciferase
measurement. The lysis buffer must be purchased separately.
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