Firefly luciferase luminescence assays using scintillation counters for quantitation in transfected mammalian cells |
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Authors: | V T Nguyen M Morange O Bensaude |
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Affiliation: | Département de Biologie Moléculaire, Institut Pasteur, Paris, France. |
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Abstract: | The firefly enzyme luciferase catalyzes the luminescent reaction of luciferin with ATP and oxygen. The luciferase gene has recently been cloned and proposed as a reporter gene in procaryotic and eucaryotic cells. We present here a luciferase activity assay which relies on luminescence detection using a standard scintillation counter. This technique is simple, fast, inexpensive, and still very sensitive: as little as 0.02 pg (250,000 molecules) of enzyme is readily detected. The technique is optimized for the luciferase assay in mammalian cell lysates. Thus, the luciferase gene may become a very useful tool for gene regulation studies. |
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