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The firefly luciferase reaction: applications, optimisation and standardisation Arne Lundin* BioThema AB, Strandvägen 36, S-130 54 Dalarö, Sweden      *(biothema@swipnet.se) Firefly luciferase is widely used for assays of ATP and for continuous monitoring of ATP producing or degrading reactions. The presence of ATP in all living cells is used in cytotoxicity/cell proliferation assays, rapid microbiology and hygiene control. In reporter gene technology firefly luciferase is one of the most important analytes. Reagent requirements in these diverse applications differ considerably. We have developed a series of ATP reagents with detection limits from the attomolar to the femtomolar range. However, the detection limit is not the only analytical parameter of interest. Recombinant luciferases can be developed to meet different application requirements. This development will put more emphasis on how to optimise, standardise and characterise the resulting new reagents. A stable ATP standard in liquid form is now available facilitating calibration of ATP assays. An assay of luciferase activity, which provides the same numerical value with different luminometers and at any ATP concentration well below the Km value, is described. A wide-spread use of these tools would be a step towards standardised ATP reagents and would facilitate comparisons of different luciferase preparations and ATP reagents. [Talk: lundin.arne.17921]

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