Enzyme Substrates and Kits for Luciferase

A variety of species have evolved the ability to emit light biochemically. Among those that have found the most use in molecular biology applications are the North American firefly photinus pyralis, the marine copepod Gaussia princeps and the bacterium Vibrio harveyi.

The firefly luciferase gene (luc) has been cloned into a variety of species, becoming a popular marker gene for plants, becteria and mammalian cells. Measurement of transciptional regulation by coupling the expression of a reporter gene is widely used to monitor a variety of physiological events including the action of specific receptor elements on gene expression, signal transduction, expression of transcription factors, protein-protein interactions, viral infection and processing. The firefly luciferase gene encodes a functional protein immediately upon transcription, and an assay for the resulting light production is highly sensitive (very low background), rapid, and reliable, making luciferase a popular choice for reporter cloning experiments. These assays are also well suited to automation and high-throughput applications.

The substrate, coelenterazine is common to a number of marine bioluminescent reactions, including those from Renilla, Gaussia, Aequorea and Watesenia. In some of these reactions it is utilized as a simple substrate being catalytically turned over in the bioluminescent reaction, while in others, such as in the photoprotein systems of Mneiopsis, it is incorporated as part of the photoprotein. The substrate h-coelenterate luciferin is a coelenterazine luciferin analog wherein one of the phenolic groups is replaced with a proton. This substrate exhibits a high recharging efficiency and even higher light output.

CHEMILUMINESCENT SUBSTRATES AND KITS

PRODUCT #	PRODUCT NAME	WAVELENGTH OF RELEASED LIGHT
M0739	Coelenterazine (native form)	470nm
M0237	D-Luciferin	560nm
M0626	Live Cell Luciferase Assay Kit	560nm
M0236	D-Luciferin, 6' Methyl Ether (inhibitor of firefly luciferase)	560nm
M0906	D-Luciferin, Ethyl Ester (membrane permeant analog of D-Luciferin)	560nm
M0352	6-Amino-D-Luciferin (analog of D-luciferin for use in sensitive peptidase assays)	560nm
M0802	6-FMOC-Amino-D-Luciferin	560nm
M0353	N-CBZ-b-Glycinyl-6-Amino-D-Luciferin (for use with a-chymotrypsin)	560nm

REFERENCES:

1. Methods Cell Biol 50, 425 (1995)
2. Methods Mol Biol 55, 147 (1995)
3. J Gen Microbiol 138, 1289 (1992)
4. Meth Mol Cell Biol 1, 107 (1989)
5. Methods Mol Biol 7, 237 (1991)
6. Biotechniques 7, 1116 (1989)