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Dual-Luciferase Quantitation in the LMax Microplate Luminometer (MaxLine Application Note #39)

/M value, so the ratios appear automatically in the group table. If, instead, the reduction is set to P value or M value, that set of values would appear in the group table.

If you wish to list both the firefly and renilla values in the group table, as well as the ratios, you can create additional columns and type in the desired formulas. Those formulas are summarized in the table below.


An example of a group table containing the 3 values (and other information) is shown in Figure 4. In this example the purified enzyme preparations were diluted in tandem, so the ratios are predictably the same for all of the samples (except the very lowest concentrations). In real cell-based experiments, the ratios would differ.


SUMMARY
The Lmax microplate luminometer is a sensitive and easy-to-use instrument for measuring firefly and renilla luciferase activity in a microplate format. The dual sequential measurements can be automated, and SOFTmax PRO for Lmax offers automatic calculations of the ratios of the two activities as well as additional powerful data analysis capability.


REFERENCES

1. www.promega.com

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

3. Matthews, J.C. et al. (1977) Purification and properties of Renilla reniformis luciferase. Biochemistry, 16:85.


By default, SOFTmax PRO brings only the single set of reduced data values into the group table. If you want additional data listed in the group table (e.g. more than one reading or additional secondary calculations), you must create new columns and specify the desired values by typing in custom formulas. For details, see your SOFTmax PRO Users Manual.




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