Reportergene Assays
Bioluminescent Basics:
Bioluminescence is a natural phenomenon that has evolved independently many times in a wide range of species like bacteria, unicellular algae, coelenterates, beetles, fishes and others. This bioluminescent effect is created by light production by an exergonic reaction of molecular oxygen with different substrates (luciferins) and enzymes (luciferases). The most popular and characterised luciferases are from Firefly (Photinus pyralis, Luc), the sea pansy Renilla reniformis (RLUC), Gaussia princeps (GLUC) and Vibrio fischeri (Lux-System).
The Firefly luciferase is the mostly used bioluminescent reporter and is a monomeric protein of 61kDa. The reaction can be regarded as a two step oxidation reaction to yield light (green to yellow region, 550-570nm). In first the reaction of luciferin (LH2) with ATP and Mg2+ generates inorganic pyrophosphate and an (the light emitter) producing CO2, A intermediate luciferyl-adenylate (LH2-AMP). The second step is the oxidation and decarboxylation of LH2-AMP to oxyluciferin mono-phosphate (MP) and releasse of photons.
go to Alkaline Phosphatase
go to Beta-Galactosidase-Assays
go to Beta-Glucuronidase-Assays