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Luciferase Reporter Gene Cells and Recombinant Luciferase Dilutions Studied with the CLIPR Luciferase Assay Kit and the LMax Microplate Luminometer (MaxLine Application Note #38)

ntially instantaneous cell lysis.


DYNAMIC RANGE AND SENSITIVITY
Figure 4 illustrates typical results obtained with recombinant firefly luciferasestandards. The dynamic range spans at least 6 logs.


The low end of the standard curve is shown in Figure 5. The lower limit of detection (based on amount giving a signal 3 positive SD of the blank) is approximately 2 fg/well, and the lower limit of quantitation (based on amount giving a signal 10 positive SD of the blank) is approximately 6 fg/well.


PCRE-LUC-CHOK1 CELL RESPONSE AS A FUNCTION OF CELL NUMBER
The magnitude of the luminescent signal clearly increases as the number ofcAMP-treated cells is increased (Figure 6).


SUMMARY
Molecular Devices CLIPR Luciferase Assay Kit makes the measurements easy and sensitive with a lower limit of quantitation for (recombinant enzyme) of approximately 6 fg/well. For measuring luciferase in cells, lysis and enzyme activity occur almost instantaneously in a flash-type reaction. The Lmax microplate luminometer is an excellent instrument for measurement of luciferase activity as a reporter gene in a microplate format. In addition, SOFTmax PRO for Lmax provides a powerful and convenient instrument control, data analysis and presentation package.


REFERENCES
1. DeLuca, M.A. and W.D. McElroy, (1978) in Meth. Enzymol. vol 53:, p.3


Reading with no delay may give slightly better results in view of the fact that both the flash reaction and cell lysis appear to be maximal within 2 seconds (see Figures 2 & 3).


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