Firefly luciferase luminescence assays using scintillation counters for quantitation in transfected mammalian cells.
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.
Nguyen
V T
VT
Département de Biologie Moléculaire, Institut Pasteur, Paris, France.
Morange
M
M
Bensaude
O
O
D000818:Animals; D002021:Buffers; D006863:Hydrogen-Ion Concentration; D007739:L Cells / Q000032:analysis; D008156:Luciferases / Q000032:analysis* / Q000235:genetics; D008163:Luminescent Measurements; D051379:Mice; D012588:Scintillation Counting; D014162:Transfection
D002021:Buffers; D008156:Luciferases