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Luciferase Assays

Answer: To visualize Firefly and Renilla luciferase activity, it is necessary to first lyse the cells. On the other hand, GeneCopoeia’s Secrete-Pair™ Dual Luminescence Assay system uses two secreted reporters: Gaussia luciferase (GLuc) and secreted alkaline phosphatase (SEAP). Secreted reporters can be detected in the cell medium without lysing the cells. Therefore, you can assay the same sample multiple times, which is provides greater accuracy and reproducibility than assaying multiple samples in applications such as time course assays.
Answer: “RLU” stands for “Relative Light Unit”. Luminometers do not measure light output, in terms of numbers of photons, directly.
Answer: All GeneCopoeia luciferase kits are compatible with all known luminometers.
Answer: All GeneCopoeia luciferase reagents are stable for at least 6 months if stored at -20°C.
Answer: To prevent loss of activity, it is best to avoid repeated freezing and thawing. We recommend pipetting small aliquots into new, clean, sterile tubes and placing those aliquots at -20°C. Once you have thawed one of the aliquots, discard it instead of putting it back in the freezer.
Answer: The Gaussia luciferase substrate, coelenterazine, is also the substrate for Renilla luciferase, so it is compatible with Renilla. However, coelenterazine cannot be used with Firefly (Photinus pyralis) luciferase, which instead uses luciferin as its substrate.
Answer: Unfortunately, the individual components of GeneCopoeia's luciferase kits cannot be purchased separately at this time.
Answer: Typically, between 12 and 24 hours.
Answer: Buffer GL-S, which provides greater signal stability, is most useful when assaying a relatively large number of samples. Its enhanced signal stability means that there will be little sample-to-sample variation due to signal decay. Buffer GL-H is used when maximal signal intensity is required, but the trade-off is that the signal decays more rapidly under typical laboratory conditions. Buffer GL-H is not appropriate for measuring the activities of large numbers of samples, unless sample injectors are used.
 
Answer: The Luc-Pair™ Duo-Luciferase Assay Kit 2.0 (For Enhanced Stability), Luc-Pair™ Duo-Luciferase Assay Kit HT (For High Throughput), Luc-Pair™ Firefly Luciferase HT Assay Kit, and Luc-Pair™ Renilla Luciferase HT Assay Kit (For High Throughput), which provide greater signal stability, are most useful when assaying a relatively large number of samples. Their enhanced signal stability means that there will be little sample-to-sample variation due to signal decay. Conversely, the Luc-Pair™ Duo-Luciferase HS Assay Kit (For High Sensitivity), Luc-Pair™ Firefly Luciferase HS Assay Kit (For High Sensitivity), and Luc-Pair™ Renilla Luciferase HS Assay Kit (For High Sensitivity) are used when maximal signal intensity is required, but the trade-off is that the signal decays more rapidly under typical laboratory conditions. These kits are not appropriate for measuring the activities of large numbers of samples, unless sample injectors are used.