Description
Simple Non-Homogenous Assay to Quantify Firefly Luciferase Activity
The Luciferase Assay System is an extremely sensitive and rapid reagent for quantitation of firefly luciferase. Linear results are seen over at least eight orders of magnitude of enzyme concentration, and less than 10 –20 moles of luciferase can be measured under optimal conditions. Generally, 100-fold greater sensitivity can be achieved over the chloramphenicol acetyltransferase (CAT) assay.
The Luciferase Assay Reagent generates light that is nearly constant for at least 1 minute and so the assay is compatible with measurement of luciferase using a single-tube luminometer or a multiwell plate luminometer with an auto-injector. A luminometer is preferred, but not required as the assay is adaptable to scintillation counters.
Cell Lysis Reagents used with the Luciferase Assay System
The Luciferase Assay System is a nonhomogeneous assay; the cells containing the luciferase must be lysed before reagent addition. Glo Lysis Buffer (Cat.# E2661), Cell Culture Lysis Reagent (Cat.# E1531), Passive Lysis Buffer (Cat.# E1941) and Reporter Lysis Buffer (Cat.# E3971) can be used with the Luciferase Assay System for reporter quantitation in mammalian cells. Reporter Lysis Buffer enables luciferase, CAT and β-galactosidase assays to be performed from the same cell extract. The Luciferase Assay System also can be used for quantitation in plant and bacterial cells, but only Cell Culture Lysis Reagent is suitable for these applications.
In some kits the lysis buffer is included, and in others it must be purchased separately (see the Specifications section of this page to view assay components).
References
Wood, K.V. (1991) In: Bioluminescence & Chemiluminescence: Current Status, eds. P. Stanley and L. Kricka, John Wiley & Sons, Ltd., Chichester, 543.
Seliger, H.H. and McElroy, W.D. (1960) Arch. Biochem. Biophys. 88 , 136.
Wood, K.V. et al. (1984) Biochem. Biophys. Res. Comm. 124 , 592–6.
de Wet, J.R. et al. (1985) Proc. Natl. Acad. Sci. USA 82 , 7870–3.
Alam, J. and Cook, J.L. (1990) Anal. Biochem. 188 , 245–54.
Schenborn, E. and Goiffon, V. (1993) Promega Notes 41 , 11–3.
Jones, D.P. et al. (1995) Promega Notes 54 , 20–5.
